I believe that life is chaotic, a jumble of accidents, ambitions, misconceptions, bold intentions, lazy happenstances, and unintended consequences, yet I also believe that there are connections that illuminate our world, revealing its endless mystery and wonder.
—David Maraniss, American author and journalist1
SOMETIME AROUND THE middle of 1961, an electrical freezer failed at the Wistar Institute. In it, according to Hayflick, were his frozen stocks of all twenty-five WI cell lines. The accident was discovered too late, and he lost all of the cells. One year earlier this would have been a major setback. By this point, however, it was mainly an annoyance. The cells had already yielded up their data; Moorhead had captured their chromosomes, in all their glorious normality, on film; and the paper had been written up, submitted, and accepted. It would appear in print in Experimental Cell Research in December 1961.2 And Hayflick, if he knew anything by now, knew just how to make replacements. So this wasn’t the end of the world. It was more like the beginning.
Hayflick had become firmly convinced of the potential of normal human fetal fibroblasts to improve vaccine making. In his view they were far superior to the expensive, sometimes-infected monkey kidney cells then being used to make polio vaccine—the only vaccine against a common viral illness that was then available. What was more, the human diploid cells could be infected with viruses for which vaccines didn’t yet exist or were in early development: common viruses including measles, chicken pox, and adenovirus, which frequently caused respiratory infections. Why not use the diploid cells to develop these new vaccines? Perhaps too his fetal fibroblasts could be used to improve on the two rabies vaccines that were then in use—one made from the pulverized brains and spinal cords of rabies-infected rabbits and the other from duck embryos. The first could have serious or fatal side effects. The second was sometimes ineffective.fn1
Hayflick determined that he would make a human cell strain that, if all went well, might become a gold standard for vaccine manufacturers. Not a strain launched, as he had launched the first twenty-five WI strains, with an exploring scientist’s tentativeness and curiosity, but one created with an eye to the future. A human diploid cell strain that vaccine manufacturers would embrace because of its pedigree: it would be free of viruses, free of cancer, and available in such quantities that running out of it would never be a problem.
To his mind such a gold-standard cell strain would have a huge advantage over, for instance, monkey kidney cells—which came from endless new pairs of monkey kidneys, each with its own risk of harboring some unwanted virus. This was because the monkey kidney cells used to grow the polio vaccine virus were used just once, when they were freshly harvested from the kidneys of a newly slaughtered monkey. They were not allowed to divide repeatedly into ever-expanding quantities in lab bottles because regulators feared that the cells might turn cancerous with repeated divisions. As a result, tens of thousands of animals were imported and slaughtered each year to make polio vaccine.3
By contrast, a human cell strain derived from just one pair of fetal lungs and then allowed to replicate could be established as clean and safe at the beginning and then used without worry going forward. Not to mention that it would be far less expensive to provide: one pair of fetal lungs would be all that was needed. The costly obtaining and sacrificing of unending numbers of monkeys could stop.
Hayflick didn’t relish the prospect of going back to the gynecological surgeons at the Hospital of the University of Pennsylvania to ask for a twentieth fetus. For one thing, they were surgeons and had a limited interest in the esoteric projects of a junior biologist from the research institute across the street. For another, Hayflick knew that he was now going to need more than a fetus. He was going to need a family history of the parents of that fetus. It would have to be a clean history that would reassure vaccine makers there were no infectious diseases or cancer lurking in either parent—conditions that would “scare the hell” out of them, as Hayflick recalled in a 2012 interview.4 For this kind of cooperation he would need, on the upstream end of the abortion, a scientist who understood vaccine making and who therefore understood the importance of his project.
Sven Gard was a tall, solemn, soft-spoken Swede who had spent eight months on sabbatical at the Wistar Institute beginning in January 1959. He had occupied a lab across the hall from Hayflick’s and one door down. There he worked with the lights half dimmed—perhaps, Hayflick thought, because the low light made a man used to long northern winters feel at home.
The gray-eyed Gard was brilliant and renowned. Fifty-three years old, he was a father figure in virology who inspired both fear and adoration in his students. He was a power player of the sort that Koprowski regularly recruited to the institute. As chairman of virology at the famous Karolinska Institute in Stockholm, which awards the Nobel Prize in physiology or medicine, he was regularly a member of the committee that decided the winners of the coveted award. He had been instrumental in seeing that the prize went to Enders and his colleagues in 1954, for their discovery that polio virus would grow in many kinds of human cells and not only nerve cells, which opened up the quest for a polio vaccine.5
Gard himself had played a major role in his country’s intense drive to develop its own polio vaccine—an urgent goal because Sweden had a far-flung population and corresponding reservoirs of people who had never been exposed to the virus and had therefore never developed antibodies. That made the country particularly vulnerable to polio epidemics. In the 1950s Sweden had the highest number of cases per capita anywhere.6
Gard was a big believer in making human vaccines in human cells. He had been inspired by the idea during a 1952 visit to the Enders lab in Boston, where he saw the polio virus being grown in human cells. On his return to Sweden, where abortion had been legal since 1938, he began using cells from human fetuses obtained from hospitals in Stockholm to develop a Swedish polio vaccine.7 Unlike Hayflick six years later, Gard’s team of virologists didn’t try to coax the human fetal cells to replicate over and over again in lab dishes. Instead they used the fetal skin and muscle cells just once, infecting them with polio and then harvesting the virus-laden fluid that bathed the cells.
The skin and muscle were the only organs sufficiently big to give them enough cells to work with. But nonetheless they found that their efforts yielded ten times less vaccine virus than other scientists were producing using monkey kidney cells. While the Gard team did run one human trial, vaccinating two thousand children with the fetal cell–grown polio vaccine, it became clear that they could not generate enough of it to vaccinate a population of seven million people.8 Gard eventually bowed to the inevitable and used monkey kidney cells to make the Swedish vaccine.
Why didn’t Gard and his team simply grow huge numbers of the human fetal cells, coaxing them to replicate in lab dishes? “We never thought of it,” says Erik Lycke, who was a twenty-seven-year-old MD/PhD student working for Gard in 1953. “I don’t think anyone but Hayflick did.”9
The Swedes launched their monkey cell–produced polio vaccine in 1957. By 1964 the disease would be virtually eliminated in Sweden. In January 1959 Gard, who would later say he had been “breathing” polio research night and day, at last found time to respond to Koprowski’s invitation to take a sabbatical at the Wistar Institute. There he met, among others, Hayflick.
Hayflick recalled that Gard, who died in 1998, overheard him griping about getting fetuses from the gynecologic surgeons at HUP—the hassle of it and their lack of understanding of his purposes—and volunteered that if Hayflick should need fetuses in the future, it was easy for him to get hold of them back in Sweden.
Possibly as early as 1959, Hayflick began to take advantage of Gard’s offer, receiving occasional fetuses or fetal organs from Sweden. The time required in transit was not an obstacle; Hayflick had discovered soon after he began working with fetuses that living fetal tissue could be kept for five days at room temperature without dying. Minced tissue floating in growth medium lasted even longer: up to three weeks.10 Now, in 1961, confronted with the demise of his first twenty-five fetal cell lines, Hayflick turned again to the solemn Swede.
Eva Herrström had been working with Sven Gard since 1952 in the Karolinska Institute’s virology department, which was housed on the expansive grounds of Sweden’s National Bacteriological Laboratory. She had practically grown up there, where her father, Josua Tillgren, a prominent Stockholm physician, had hired her as an assistant lab technician in the summer of 1943, when she was just seventeen. She never left, and by the mid-1950s she had risen to become Gard’s top technician, a position she still held in 1961.
Gard’s lab wasn’t in the elegant main building designed by the famous modernist architect Gunnar Asplund but in a small, two-story yellow-brick outbuilding where it occupied the main wing of the second floor. The temporary-looking building was nicknamed the Monkey House because the other wing housed African green monkeys used to safety-test polio vaccine. The communal freezer was just outside the windowed door into the monkey wing; whenever someone deposited a sample in it, the monkeys shook their cages, raising a ruckus.
Humble digs or not, Herrström liked her work as Gard’s chief lab technician. He took an interest in all of his staff, inviting her and the other technicians when he presented lectures on scientific advances and crediting everybody for his or her contribution. Still, there was no mistaking who was in charge. There was little or no frivolity on the job, no happy hours, no practical jokes or undue familiarity. She knew that she would, until her dying day, call Gard “Professor.”
On this particular morning—April 24, 1961—Herrström climbed the metal stairs that ran up the outside of the building to the second floor to learn that a fetus would be arriving and that she needed to prepare its lungs for shipment to the United States.
When she had first worked for Gard almost a decade earlier, he had been trying to make polio vaccine with human cells. Then, Herrström had worked plenty with human fetuses, even learning how to expertly drain amniotic fluid that was used in cell-nurturing medium from the intact pregnant uteruses of cows that arrived regularly from the Stockholm slaughterhouse. But once Sweden moved to using monkey cells to develop the vaccine, that work with human fetuses had gone away—until lately, when Gard had begun asking her to prepare tissue for shipment to an American institute in Philadelphia.
Later, after she had donned a white gown and a car had delivered a tiny bundle wrapped in green surgical cloth, Herrström headed for one of the sterile rooms in the middle of the floor. She washed her hands in disinfectant at the sink under the window, laid out her instruments, and sat down at the shiny linoleum table that was the lone piece of furniture in the room. She unwrapped the bundle.
It really was incredibly beautiful, this little fetus, with everything already in place. With this task she was being given a privileged glimpse into the creation of life. It helped to remember this as she picked up a scalpel. And it helped that she came from a family of physicians. You got used to it. You turned the tragedy around. You said to yourself that at least in this case, something life-giving might emerge from death.11 What shape that particular good might take in this instance Herrström didn’t know. But if Professor Gard said that scientists in Philadelphia needed fetal lungs, her job was to make sure they got them.
At home that night Herrström made a dinner of sliced reindeer meat and repaired her winter gloves. Before turning in at 9:30 p.m., she wrote in her diary: “Work 8.30-[5 pm]. Sent tissue to USA. Stressful.”12
The lungs that Herrström had earlier that day placed in medium in a small test tube, and the tube that she had then placed in a thermos, and the thermos that had then been packed in a box, on wet ice, were well on their way by then to Philadelphia.
It’s probable, although not certain, that the lungs that Herrström removed from that fetus on that spring day went on to become the next cell strain that Hayflick created. (By this point Hayflick was working only with lungs because they were readily dissected and their fibroblasts seemed particularly hardy in culture.) It’s also possible that those lungs didn’t work out. Perhaps delays caused them to die in transit, or perhaps they were inadvertently infected. Because scarcely one week later Herrström recorded in her diary that she had shipped more tissue, from a new fetus, to Philadelphia: “Tissue to the USA at 12 (noon). Took all morning to prepare.”13
What is certain is that by the autumn of 1961 Hayflick had launched his twenty-sixth diploid cell strain, from the lungs of a male fetus aborted sixteen weeks into pregnancy and sent to Philadelphia from Gard’s lab in Sweden. By that October he had taken time-lapse photos of the new WI-26 cells being attacked by polio virus in a lab dish.14
Hayflick’s timing in launching the new fetal cell strain was near perfect. He produced and froze what he thought would be plenty of the fibroblasts—about two hundred ampules of them, each containing up to two million cells—shortly before the December publication of “The Serial Cultivation of Human Diploid Cell Strains,” the paper that announced to the world his discovery of the Hayflick limit and his cultivation of those first twenty-five normal fetal cell strains. Readers, of course, didn’t know that those cells had since died in the freezer failure.
Demand for his human fetal cells soared when the landmark paper was published. With the dozens of viruses that Hayflick had demonstrated to infect the cells, virologists were keen to get hold of them for experiments on the nature of these viral diseases. Companies hoping to launch viral vaccines wanted them. Basic biologists too sought them for all manner of study on the workings and behavior of normal cells in lab dishes. With the new WI-26 cells on hand, Hayflick was ready.
Soon he was handing out ampules of WI-26 left and right to biologists, virologists, and companies aiming to create vaccines against measles, adenovirus, and polio. But before he knew it, to his chagrin, Hayflick began running out of ampules—an embarrassing fact, since he had been determined to make a plentiful supply. What Hayflick hadn’t reckoned on was the demand for the cells. Had he seen it coming, he might have been more conservative, thawing one ampule at a time and expanding its contents through several generations in the incubation room, then sending out the much-more-numerous resulting cells. Giving away the ampules themselves was like giving away his seed corn.
“As a consequence of the unanticipated and unprecedented demand” for WI-26 cells, he would write in the Wistar’s next biennial report, “depletion” of his stocks had occurred.15
He needed to begin again—again. He needed to create a human fetal cell strain that would outlast the current, seemingly bottomless demand, for this new kind of normal, diploid cell. Fortunately, Gard continued to be good on his word. This time Hayflick asked Gard for the lungs of a female fetus, to ensure that if any of the male WI-26 cells he had launched into labs around the world became mistakenly mixed with the new cells during an experiment, the problem could be made quickly apparent by looking at the cells on a microscope slide: in a female cell strain, there should be no Y chromosomes present.
It wasn’t easy for Gard to deliver just the right fetus quickly. It had to be female. It had to be large enough—from a pregnancy three to four months along—to have readily dissected lungs that would yield enough tissue. And it had to be from a woman without health problems in her present or past. And so Hayflick waited. At last the lungs of a female fetus arrived from Sweden. Continuing with his numerical order, Hayflick named this next female lung-cell strain WI-27. Shortly after launching it, he handed a sample of the cells to Moorhead. He examined the cells and reported back that there was an abnormality in the WI-27 chromosomes. It was probably inconsequential, but “probably” was nowhere near good enough for vaccine makers, wary as they were of using cells that might be cancers in waiting. The strain was useless for vaccine-making purposes. Hayflick would have to go back to Gard once again.
At this point another scientist might have thrown up his hands and moved on to something else. The groundbreaking work deriving these normal human fetal cell strains was done. Hayflick’s methods were now published for all to see. What was more, he had launched a goodly supply of WI-26 cells into academic and commercial labs. Another scientist might have told colleagues and companies that if they wanted more normal cells, they could make their own. All it involved was engineering and practice. This line of thinking, however, was completely foreign to the dogged, determined, ambitious man who was Leonard Hayflick, who was set on delivering a lasting human diploid cell strain to the world.
Besides which, he was now working under a new obligation. In February 1962, as Hayflick was watching his supply of WI-26 cells rapidly dwindle, the National Cancer Institute, part of the National Institutes of Health, the U.S. government’s medical research agency, responding to the keen and obvious demand for Hayflick’s new breed of cells, entered into a contract with the Wistar Institute. Under it the Wistar—meaning, in practice, Hayflick—committed “to produce, characterize, and store human diploid cell strains” and to distribute them to all qualified investigators.16 From 1962 the Wistar began receiving from the National Cancer Institute annual contract payments of at least $120, 000—nearly $1 million in 2016 dollars and about 10 percent of the Wistar Institute’s income from grants and contracts.17
Koprowski’s institute did need to make an initial outlay: In June 1962 the institute’s board of managers approved the Wistar director’s request for $8,000 to retrofit Hayflick’s “new Diploid Cell Laboratory” with a power line, extra air-conditioning, and plumbing that the NIH contract didn’t cover.18 Given the size of the contract, they must have felt that the payoff would more than justify that investment.
The language of the contract may seem dull, but it will become important. It stated in part that when the contract was terminated, “the contractor agrees to transfer title and deliver to the Government, in the manner, at the time and to the extent, if any, directed by the Contracting Officer, all data, information and material which has been developed by the Contractor in connection with the work under this contract.”19 In other words, while Hayflick would be the developer of new diploid cells under the contract, the government would own them, and when the contract was up, he would need to transfer any cells he had developed to the National Cancer Institute or place them wherever the agency directed him to put them. (In the case of cells, privately held cell banks are often used as cell custodians.)
And so, working under this new contract obligation, Hayflick, after the failure of WI-27, asked Gard for another female fetus. And once again he waited.
Hayflick made one more decision in the wake of the WI-27 disappointment. With the next human diploid cell strain he was going to change the numbering, because some scientists seemed to be carelessly confusing the numbers of earlier diploid cell strains. In order that going forward from WI-27 there would be no more such confusion, he would raise the “2” to a “3” and the “7” to an “8.” The next cell strain he created would be called WI-38.
On the warm, sunny morning of June 7, 1962, Eva Ernholm prepared for her job as an obstetrician/gynecologist at the Women’s Clinic in a hospital near Stockholm. One of few women in Swedish medicine in 1962, the thirty-seven-year-old Ernholm was an adventurer. As a medical student driven partly by an interest in Eastern religions, she had worked in a U.S. Eighth Army MASH unit in the spring of 1951, during heavy action in the Korean War, and returned to Korea with the Swedish Red Cross in 1953. She came back from her journeys bearing photos of herself bent over patients in a field hospital operating room and in fatigues surrounded by orphans, looking like she was having the time of her life.20
Ernholm was a blunt woman who knew her own mind and didn’t hesitate to speak it. Whether people liked her or her views—and they inevitably felt one way or the other—was a matter of indifference to her. She was impulsive and decisive and passionate. And, like Hayflick, she was not someone who was turned aside easily. An accomplished pianist, she had insisted on using a construction elevator to lift her grand piano to the penthouse apartment where she had taken up residence one year earlier.
Ernholm had abandoned an early interest in neurology to study obstetrics and gynecology and was a newly trained specialist when she took her hospital job in 1961. At the Women’s Clinic she shared a heavy workload with one other junior doctor, monitoring pregnant women, delivering babies, cutting out uterine growths, and tying women’s tubes when they requested it. She also performed abortions.
Ernholm did not do this lightly. The following year, 1963, she would tell a newspaper reporter, “As a matter of principle, I am against all abortions that are not performed for medical reasons.” But, she added, “evidently there are situations when the social circumstances are so valid they don’t leave you any choice.”21
On this lovely June day Ernholm was confronting such circumstances, in a woman in her thirties who throughout this book will be called Mrs. X. Her medical history was uneventful, apart from childhood bouts with whooping cough, measles, and scarlet fever. Really, her only problem was that her last menstrual period had been in late January. Mrs. X explained that she already had several young children, and that her run-down husband, a working man, was often out of town for his job. When he was home, he wasn’t much use: he was an immature alcoholic who had done time in prison.
Ernholm took out her stethoscope and pressed it to Mrs. X’s chest and back. Her patient’s heart and lungs were clear, as they would need to be for the operation. Ernholm laid a hand on the woman’s soft abdomen. It was painless, Mrs. X confirmed, when Ernholm pressed it. Now came the stirrups and the hard metal of the speculum. It couldn’t be helped; one had to be sure that the patient was pregnant. Ernholm looked and then noted in Mrs. X’s record that the cervix, the mouth of the womb, was bluish—a telling sign of pregnancy. By feel and by sight, Ernholm judged that she was sixteen or seventeen weeks pregnant. She recorded this and wrote: “Indication for abortion: General weakness.” Then she added the details of Mrs. X’s home situation.
Sweden, where abortion had been a capital crime one hundred years earlier, had legalized abortion in 1938. The law enacted that year stipulated that women could have abortions in three situations: in cases of rape or incest; if delivering the child would cause “sickness” or “weakness” that seriously endangered the mother’s life or health; and for “eugenic” reasons, meaning that the mother or father was likely to transfer a serious hereditary disease to the child. (In this case the woman had to agree to be sterilized at the time of the abortion.) In 1946 the law was liberalized slightly to include “expected weakness” of the mother as one of the conditions that made an abortion permissible—the assessment that, given the mother’s living situation and circumstances, her health could seriously deteriorate if she was to bear and raise the child.22
In practice, however, getting an abortion in Sweden in 1962 was far easier said than done. A woman who sought to end a pregnancy had two choices. She could apply to the Medicinalstyrelsen—the Royal Medical Board—which regulated abortions for the government from an imposing nineteenth-century building in Stockholm. (It memorialized Swedish King Oscar I, an honorary member of the Swedish Academy of Sciences and a prolific father who sired eight children—three of them by mistresses.)
Alternatively, a woman could try to convince two doctors that she needed to end her pregnancy. One of the doctors was often a psychiatrist. The other was the surgeon who would perform the procedure. Most Swedish doctors opposed abortion and refused to help women get them, leaving thousands of women’s applications to grind through the slow-turning gears at the government’s Royal Medical Board, often pushing abortions well into the second trimester.
Mrs. X was seeking to end her pregnancy at just about the worst time possible for a woman in Sweden who wanted an abortion. In 1951 the Swedish Medical Association had adopted ethical rule IV, which said that the physician “should consider his duty to protect and preserve human life from its implantation in the mother’s womb.”23 The next year the chairman of the medical association, in the midst of a debate about whether to rescind the 1946 change that liberalized the law, said that abortion was in the same category as child murder.24 Swedish abortion rates then fell markedly, reached a low point in 1960, and scarcely budged in the next two years.
The day after Ernholm examined her, Mrs. X was wheeled into an operating room, where Ernholm performed what was called a “minor Caesarean section.” She cut through Mrs. X’s abdominal wall, carefully dissected the bladder free from where it lay high on the uterus, and cut through the wall of the uterus. She removed the fetus and the placenta, being careful not to leave any tissue behind. “The cavern was cleaned. Suture of the uterus in stages,” Ernholm wrote in the operative report. The fetus, she noted, “was 20 cm. long and female.”
That fetus was wrapped in a sterile green cloth, handed to an aide, and taken to a car for transport to the virology department of the Karolinska Institute.
A few days later, on a gray, drizzly morning in mid-June, Hayflick sat down in one of the tiny “sterile” rooms in his lab. Following a routine that was now deeply familiar, he dipped a pair of tweezers in alcohol, flamed them in a Bunsen burner, waited for them to cool, and then lifted two small, purplish chunks of tissue from where they floated in a glass bottle of clear pink solution. He laid them on a petri dish. Using a pair of scalpels held at right angles, working them like an improvised scissors, he minced the lungs into innumerable pinhead-size pieces. He deposited them in a flask, where a trypsin solution would break down the connective tissue holding the cells together, releasing millions and millions of individual cells.
Later he poured that mixture into several small glass tubes, stoppered them, and loaded them into a centrifuge, a round machine that sat on a pedestal with wheels and could be moved here and there around the lab. He turned it on and the tubes began spinning so fast that they flew out at an angle horizontal to the machine. After twenty minutes or so, the cells, being heavier than the fluid, sank to the bottom of each tube as an off-white pellet.
He turned off the machine, recovered the tubes, and, back in the sterile room, repeatedly blasted the pellet in each tube with medium, using a glass pipette with a cotton stopper and the power of his lungs. Eventually the cells came loose from one another. He sucked them up in the pipette, transferring them bit by bit into a big glass bottle. Moving quickly now so the cells wouldn’t stick, he poured the mix of cells and nutrient solution into several small glass bottles. He laid these carefully in the incubation room.
Some days later, after the cells had established themselves in the bottles, Hayflick handed a sample of them to Moorhead. This time the news that came back from his friend and colleague was good. Moorhead told him that the WI-38 cells’ chromosomes appeared entirely normal.
Hayflick knew that if he could freeze a large enough quantity of WI-38 cells, he could provide vaccine makers with enough cells to make vaccines for decades to come. How was this possible, if the cells were mortal and would, sooner or later, die in their bottles? Hayflick had shown how the math worked in the landmark 1961 paper. It was all due to the extraordinary power of exponential growth.
Suppose, Hayflick wrote in that paper, you began with just one small glass Blake bottle. It was rectangular and roughly pint-sized, its larger, flat side measuring a mere 5.5 inches by not quite 3 inches. Such a bottle held roughly ten million cells when those cells had grown to confluence on its floor—really, on its side, because the bottles lay on their flat sides while they incubated. If at this point you split these newly planted cells into two bottles, and split the bottles again when the floors of those two bottles were covered, yielding four bottles; and if you then kept splitting the bottles in this way when the cells reached confluence, until the original cell population had doubled fifty times—roughly the Hayflick limit—you would produce, he calculated, 1022 cells, or 10 sextillion cells. Knowing as he did that 14.2 billion wet cells weigh about an ounce, Hayflick also calculated that the cells in that one original bottle would therefore produce twenty million tonnes of cells.25
Admittedly, this was a theoretical maximum. Real life wasn’t nearly so neat. Sometimes Blake bottles got contaminated. Sometimes the ampules in which he froze the cells cracked or, worse, exploded during thawing because liquid nitrogen had leaked into microscopic holes in their closures while they were frozen. (Hayflick took to wearing goggles when thawing ampules.) Sometimes cells got lost during shipping. And sometimes, frankly, they got thrown down the drain when there were leftovers after shipping them out to scientists.
On the other hand, there was room for a few accidents and a little waste when working with a potential ten sextillion cells—a number that’s so large it’s difficult to grasp. One way to think about it is this: the freshly harvested WI-38 cells covering the floor of just one of Hayflick’s pint-size Blake bottles, expanded until they have doubled roughly twenty times, would produce 87,000 times more vaccine than is made by a typical vaccine-making company, setting out today to make one year’s worth of a typical childhood vaccine that it will ship to more than forty countries.26
The point Hayflick was making with his calculation in the 1961 paper was that a sufficient supply of cells, frozen and thawed when needed, bit by bit, would produce all the cells that the world would need for the foreseeable future. Using his method, he wrote, “one could have cells available at any given time and in almost limitless numbers.”27
That summer, change was coming to America. As Hayflick cut the WI-38 lungs into minuscule pieces, the New Yorker published the first excerpt of Rachel Carson’s classic Silent Spring, launching the modern environmental movement. As the cells first reached confluence late in June, the Supreme Court ruled that voluntary prayer in public schools violated the constitutional separation of church and state. When President Kennedy, speaking at Philadelphia’s Independence Hall on a lovely, sunny Fourth of July, praised American democracy for encouraging dissent, three miles away the cells divided luxuriantly in thirty-six-degree heat. And as Hayflick set about creating a supply of frozen WI-38 cells that he intended to last for the foreseeable future, women’s constrained access to abortion landed in the headlines, in the person of an actress in Phoenix named Sherri Finkbine. Finkbine, a mother of four and a host on the children’s television show Romper Room, had taken the drug thalidomide to combat morning sickness early in her fifth pregnancy, unaware that it deformed fetuses. She could not get an abortion in Arizona, where state law allowed abortions only if the life of the mother was in danger.28 She ended up flying to Sweden to terminate her pregnancy, the press following her every step of the way.
In deciding when to freeze the WI-38 cells, Hayflick had to strike a balance. He wanted to produce enough cells to fill plenty of ampules for future needs. He was keenly aware that two hundred ampules of WI-26 had not been enough. On the other hand, he didn’t want the cells to get too old—to divide too many times—before he froze them. Vaccine makers wanted youthful cells that they could expand through many more divisions before they reached the end of their usefulness. They were also wary of older cells because each cell division increased the chance of chromosomal aberrations and thus, they feared, of the cells becoming cancerous.
And in fact one year after Hayflick launched WI-38, it would emerge that the chromosomes in WI-38 cells developed spontaneous abnormalities when the cells aged beyond forty divisions. These were not cancerous changes, according to the paper published in the Proceedings of the National Academy of Sciences by Moorhead and Eero Saksela, a young Finnish scientist then working at the Wistar Institute. “On no occasion,” they wrote, did any of the cells with altered chromosomes develop abnormal shapes or start to divide abnormally quickly or open-endedly, all classic signs of cancer.29
But the abnormalities that Saksela and Moorhead documented would push vaccine makers and regulators alike to steer clear of WI-38 cells at high population doubling levels. By the late 1960s companies in the United Kingdom, one of the first countries to embrace human diploid cells for making vaccines, were beginning vaccine-making campaigns with cells no older than the thirtieth population doubling level.30
Hayflick decided to freeze the cells when they had been split into new bottles eight times. Eighth passage cells—so called because they had been “passed” into new bottles eight times—would be plenty youthful, and there would be plenty of them. Just one small Blake bottle of cells, split eight times, produced 256 bottles, each containing roughly ten million cells at confluence. And Hayflick typically placed the cells from a pair of lungs into four small Blake bottles at the outset.
Some bottles, Hayflick conjectured in 2013, were probably lost to contamination, perhaps early in the splitting process, eating into the total number of bottles available when the time came to freeze the cells. At any rate, there is no record of exactly how many bottles accumulated before Hayflick gave the order that the cells in them should be distributed into tiny ampules and frozen. What is certain is that the task at hand was huge.
Hayflick was not present in his lab on the day in late July 1962 that a number of Wistar technicians, some of them borrowed from other labs, assembled to do the job. Hayflick had traveled two weeks earlier to a World Health Organization meeting being held in Geneva to discuss the potential vaccine-making uses of his human diploid cell strains. Possibly he was still on the road.
The crew of lab techs faced long hours of monotonous work. They had first to loosen the cells from where they lay in single, sticky layers on the bottoms of the Blake bottles, using lung power and the pipette technology invented by Louis Pasteur nearly one hundred years earlier. They sucked up a mix of culture medium and cells, then blasted the fluid back into the bottle and repeated this over and over again, until the fluid turned milky white with floating cells.
Then they went to work with syringes, sucking up tiny portions of the fluid and injecting these into steam-sterilized gas ampules. It was delicate work. Each little wine bottle–shaped vial was about two inches tall and had an open neck roughly one sixteenth of an inch wide. Once loaded with cells, it needed sealing. The workers—some with more finesse than others—sealed the neck of each ampule by melting it with a pass through the flame of a Bunsen burner. Using tweezers, they pulled on the string of melting glass to work it into a blunt closure—all the while trying not to kill the cells in the ampule with too much heat. They were executing what would, with hindsight, become a critically important operation: getting the cells into the ampules without letting bacteria contaminate them.
Sterilizing procedure was nowhere remotely as good in the early 1960s as it is today, in large part because technologies now taken for granted didn’t exist. For instance, today’s ubiquitous laminar flow hoods, which prevent microbial contamination of the air over a scientist’s work space, were just being developed. The ultraviolet lights then used at night had serious limits: the light they emitted killed organisms on surfaces but not in things—like cell cultures.
True, penicillin and other antibiotics were in wide use by 1962, and many labs used them liberally to protect cell cultures from bacterial contamination. But, Hayflick says, vaccine manufacturers were wary of antibiotics because of their potential for provoking allergic reactions in vaccinees. He decided to take the calculated risk of not using antibiotics. That decision would come back to bite him.
By the end of the day on July 31, 1962, the Wistar work crew had produced more than eight hundred ampules of WI-38. Each tiny vial held 1.5 million to 2 million cells. They were placed in a big, communal dry ice chest. A couple of months later Hayflick transferred them to more permanent digs in a liquid nitrogen tank in the Wistar basement. There they remained, tucked away at –160 degrees Celsius.31
In October of 1962 a seasoned cell culturist named Robert Stevenson paid Hayflick a visit at the Wistar. The straight-shooting Stevenson was Hayflick’s project officer at the National Cancer Institute, charged with overseeing the smooth and proper execution of the contract under which Hayflick was producing, storing, and distributing human diploid cell strains. Perhaps Stevenson learned during this visit that Hayflick had, ten days earlier, handed one hundred ampules of the new WI-38 cells to a visitor, Frank Perkins of Britain’s Medical Research Council, the UK’s top vaccine regulator. Perkins had taken them back to London on a transatlantic flight.32
On October 18, 1962, two days after he visited Hayflick’s Wistar operation, Stevenson wrote a memo summarizing his visit and noting that he had made it clear to Hayflick that the cells were U.S. government property.
While the cells, he wrote, couldn’t be subcontracted out to a non-U.S.-government agency for distribution, the Medical Research Council, or MRC, could function as a “distribution depot.”33 This the British agency promptly became, shipping the cells to scientists in Berlin, Madrid, Milan, Tehran, and Uppsala.34
At the end of the eventful year of 1962, Hayflick filed a progress report to Stevenson at the National Cancer Institute—a routine report owed to the government agency under the contract. It included a section entitled “Characterization of a New Human Diploid Cell Strain WI-38.”35
As soon as the WI-38 ampules were first frozen, Hayflick went back to Sven Gard in Stockholm for the documentation that he knew that regulators would require. For the purposes of the Division of Biologics Standards, the unit at the National Institutes of Health that in those days licensed new vaccines, Hayflick’s word would not be enough. He needed papers proving that Mrs. X, the mother of the WI-38 fetus, was as healthy as a horse and that no cancers, hereditary diseases, or infections were lurking in her or in the fetus’s father.
Getting that information was a delicate task. Two months after the abortion, Mrs. X had no idea that her fetus had ended up anywhere but in a medical waste incinerator. In 1962 in Sweden, as in the United States, tissue from aborted fetuses was routinely used by scientists without the knowledge of the women who had the abortions. The stern Gard, aged fifty-six and childless, was not inclined to take up this job. He delegated it to someone he may have considered better suited to approach Mrs. X: a thirty-five-year-old physician named Margareta Böttiger. Böttiger was earning her PhD in his lab while running human studies monitoring the effects of the Swedish polio vaccine.
A dark-haired beauty with an oval face who hailed from a storied and powerful Swedish family, Böttiger was reserved, mannerly, and unthreatening. Like the gynecologist Ernholm, she confronted a medical profession in which just 14 percent of physicians were women. Unlike Ernholm, she had small children—her girls were three and six years old that August—and a physician husband who did no housework. She had survived by embracing the regular hours of a scientist in Gard’s lab, rather than the pediatrics she had trained for immediately after medical school. Paying her invaluable nanny, Harriet, consumed almost her entire salary—but it never occurred to her to give up the work she loved.
The task at hand didn’t quite fit that description. But Gard had asked her to do it, and she was not one to buck authority. Böttiger telephoned Mrs. X’s primary-care doctor. Working with that doctor and the doctor’s nurse, she pieced together as much as she could and got hold of the operative record from the hospital where the abortion was performed. The whole enterprise took some doing, and it wasn’t until more than a year later—October 1963—that she wrote to Hayflick, attaching Mrs. X’s medical record.
It cataloged her childhood bouts with measles and scarlet fever and whooping cough; her freedom from other infectious diseases since then; her several healthy children; and the absence of known hereditary diseases or tumors in her family.
In her accompanying cover letter Böttiger wrote that she believed Mrs. X to be perfectly fine. But, she added, the father appeared to be subpar mentally. Böttiger also warned Hayflick, who had apparently asked for blood samples from the couple, that when Mr. X got back from his out-of-town labor, it might be tough to get his blood drawn. She did not elaborate on why.
Asked about this in 2015, Hayflick could not recall why he would have requested a blood sample from Mr. X. He added that Mr. X’s reported mental deficiency did not trouble him. Mental deficiency, he wrote, is not infectious and would not have been relevant to the cells’ safety for vaccine making.36