Lymphoblastoid Cell Line (LCL)

A lymphoblastoid cell line, or LCL, is an immortalized population of cells derived from a specific type of white blood cell called a B lymphocyte that scientists around the world began using for biomedical research in the late 1960s. By immortalized, scientists mean that the cells have been altered so they can grow and divide indefinitely, or at least for an extended period of time. That trait of LCLs makes them useful as a replenishable source of cells and the DNA contained within them. Scientists obtain LCLs by first collecting a blood sample and then exposing the B lymphocytes in the blood to Epstein-Barr virus, or EBV. EBV alters the B lymphocytes in such a way that the cells begin to multiply without restraint. Researchers began making and storing LCLs from individuals around the world in the 1960s. As of 2025, LCLs form a mainstay of biomedical research, especially in human genetics and genomics.

1. Background: Tissue Culture and Cancer Virus Research
2. Making LCLs
3. Scientific and Social Impact of LCLs

Background: Tissue Culture and Cancer Virus Research

LCLs belong to the broader history of cell culture techniques that historians trace to the work of Ross Granville Harrison, a researcher studying embryology in the US, who, in 1907, became the first to successfully culture living tissue in a medium outside of the body. Harrison, then a professor at Johns Hopkins University in Baltimore, Maryland, grew a nerve cell from a frog outside the frog’s body by implementing a technique called the hanging drop method. In that method, Harrison put a piece of embryonic frog tissue on a microscope slide cover slip, which is essentially a thin, flat sheet of glass, and coated it in frog lymph. Lymph is the fluid that circulates through lymphatic vessels and lymphoid organs such as the thymus and spleen. Harrison then inverted the cover slip over a hollow microscope slide and sealed the adjoining surfaces with wax. He found that cells from the frog tissue could live for weeks under those conditions.

In the years immediately following Harrison’s experiment, surgeon Alexis Carrel from the Rockefeller Institute for Medical Research in New York City, New York, and researcher Montrose Thomas Burrows, extended Harrison’s work, showing that it was possible to grow mammalian tissues outside the body for an extended period of time. To do so, they used Harrison’s hanging drop method but instead of using tissues from amphibians, they used tissues from mammals such as chicks. They also stated that the cultures could survive for long periods of time, but according to Milton W. Taylor, a researcher who studied viruses, that was likely not the case. Instead, Taylor explains that they might have been adding cells to the culture which created an appearance of constant growth. Carrel and Burrows published several articles on their tissue culture methods starting in 1910. Scientists also credit Carrel with developing the first form of a tissue culture flask, which is a flat container with a sealable opening on one end that allows for the cells to grow consistently.

By the mid-twentieth century, scientists had begun experimenting with using tissue culture as a means to grow viruses in the lab, in part for the purpose of making vaccines. For example, in 1940, John Franklin Enders, a researcher studying viruses in the US observed that embryonic chicken cells infected with the vaccinia virus grew constantly. Continuing into the 1940s, the Enders lab did the same with the mumps virus and then turned their attention to the poliomyelitis virus. They found that they could grow the poliomyelitis virus in human embryonic skins and muscle tissue. Before Ender and colleagues’ results, scientists had previously noted that the polio virus would grow only in neurologic tissues. The discovery that polio virus could grow in human embryonic skin and muscle tissue was, according to historian Hannah Landecker, a startling breakthrough in polio research. In meant that scientists could grow the virus in tissue culture rather than the more expensive and complicated method of growing it in lab animals. Enders, together with colleagues Thomas Huckle Weller and Frederick Chapman Robbins, received the 1954 Nobel Prize in Physiology or Medicine for their discovery. Jonas Salk, a scientist who studied viruses at the University of Pittsburgh School of Medicine in Pittsburgh, Pennsylvania, later used Enders’s method of growing polio virus, which in turn led to the first polio vaccine, in 1954.

Throughout the 1950s, scientists devised additional ways to keep human cells alive for extended periods outside the body. In 1951, scientists George Otto Gey and colleagues at The Johns Hopkins University Hospital in Baltimore, Maryland, took cancer cells from a woman named Henrietta Lacks and cultured them into what researchers refer to as HeLa cells. The HeLa cell line was one of the first immortal human cell lines. At the Johns Hopkins Hospital, surgeon Howard Wilbur Jones was treating Lacks for cervical cancer and sent the cells taken from her tumor to Gey’s laboratory. At the lab, the researchers used Lacks’s cells and cultured them into the HeLa cell line, which grew rapidly and indefinitely. However, that was all conducted without her knowledge, consent, or compensation, leading to controversy surrounding the ethics of HeLa cells. As of 2025, HeLa cells are still a mainstay of scientific research, though they do have some limitations, including the fact that their cancerous nature makes them behave differently from noncancerous cells.

In addition to the history of tissue culture, the creation of LCLs also belong to the mid-twentieth century history of research on cancer viruses such as EBV. In 1964, researchers Anthony Epstein, Yvonne Barr, and Burt Achong who were working in the UK at the time published work on a virus that can cause cancer in humans. They were among the first to note that a virus could result in cancer in humans, and researchers later dubbed the virus Epstein-Barr Virus, or EBV, after the individuals who discovered it. Then in 1967, researchers Werner and Gertrude Henle, along with their colleagues, who were working in the US published an article that reported that they could induce the continuous growth of lymphocytes, which are a type of white blood cells, by cultivating them with specially treated cells carrying EBV. The work that Gertude and Werner Henle produced allowed other scientists shortly thereafter to produce many additional LCLs from white blood cells they had obtained from human blood.

Making LCLs

As of 2025, standardized techniques exist for creating LCLs. Bryan Bolton and Nigel Spurr, researchers working in England, published a set of instructions in 1996 that researchers continue to use in the twenty-first century. First, scientists must separate lymphocytes from the other constituents of human blood. One way they do so is by using density gradient centrifugation, a technique in which a machine spins vials of blood at high speeds to separate out different components according to their density. Once scientists have separated out the lymphocytes, they then need to create an environment for EBV to infect the cells. Researchers then add together the EBV and lymphocytes with growth medium and maintain the mixture at a particular temperature for several weeks, replacing a certain amount of the liquid twice weekly with fresh growth medium. After infection, the sample of lymphocytes that researchers obtain from the previous centrifugation step will likely contain both B and T lymphocytes. Researchers require only the B lymphocytes, so as a result they typically kill off the T lymphocytes with a chemical agent called cyclosporin A, leaving only the B lymphocytes behind to grow and accumulate. Researchers can then freeze their immortalized B lymphocytes and store them indefinitely.

Scientific and Social Impact of LCLs

LCLs have several advantages over other types of immortalized cell lines. For one, according to Heidemarie Neitzel, a researcher working in Berlin, Germany, they are relatively easy to create. Scientists can make them from ordinary human blood cells that they obtain from any person through a simple blood draw, as opposed to a surgical biopsy, which is how they make other cell lines. Additionally, unlike some other cell lines, LCLs are genetically stable, which allows researchers to have access to a continuous source of DNA. For example, LCLs are useful in a variety of research projects such as studying human genome variation and using them for drug response studies.

Scientists’ use of LCLs expanded in the 1990s with the rise of human genome mapping and sequencing projects in which scientists attempted to study the DNA differences between individuals and populations. Examples of those genome projects include the Human Genetic Diversity Project, the International HapMap Project, and the 1,000 Genomes Projects. Scientists affiliated with those genome projects used LCLs as a way to preserve the DNA from the human blood samples they collected.

LCLs sidestep some of the ethical issues that have plagued the use of other human cell lines. Unlike HeLa cells, for example, LCLs typically come from the blood of individuals who knowingly participate in research. Furthermore, unlike with embryonic stem cell lines, scientists can make LCLs from adult blood cells. Embryonic stem cells come from embryos, and in order to obtain those stem cells, scientists must destroy the embryo. Certain groups have argued that the destruction of embryos for research is unethical, so researchers have looked for alternative sources of stem cells for research. Researchers have used LCLs to make induced pluripotent stem cells, which come from adult cells and so also avoid the step of destroying an embryo.

While LCLs raise fewer ethical objections than some other immortalized cell lines, they are not completely free of the potential for ethical controversy. Even if most of the LCLs in existence in biobanks around the world come from people who willingly consented to have their cells turned into a cell line, those individuals may not agree with the ways researchers use their cells lines in the future.

LCLs represent the fruits of both tissue culture research and cancer virus research spanning several decades of the twentieth century. As of 2025, LCLs remain a basic, widespread technology of biomedical research.

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