WI-38

Last updated
WI-38 cells (Left: in high density. Right: in low density) WI-38-Li-and-Tollefsbol-2011.gif
WI-38 cells (Left: in high density. Right: in low density)

WI-38 is a diploid human cell line composed of fibroblasts derived from lung tissue of a 3-month-gestation female fetus. [1] [2] The fetus came from the elective abortion of a Swedish woman in 1963. The cell line was isolated by Leonard Hayflick the same year, [3] [4] and has been used extensively in scientific research, with applications ranging from developing important theories in molecular biology and aging to the production of most human virus vaccines. [5] The uses of this cell line in human virus vaccine production is estimated to have saved the lives of millions of people. [3] [6] [7]

Contents

History

The WI-38 cell line stemmed from earlier work by Hayflick growing human cell cultures. [2]

In the early 1960s, Hayflick and his colleague Paul Moorhead at the Wistar Institute in Philadelphia, Pennsylvania discovered that when normal human cells were stored in a freezer, the cells remembered the doubling level at which they were stored and, when reconstituted, began to divide from that level to roughly 50 total doublings (for cells derived from fetal tissue). Hayflick determined that normal cells gradually experience signs of senescence as they divide, first slowing before stopping division altogether. [2] [4] This finding is the basis for the Hayflick limit, which specifies the number of times a normal human cell population will divide before cell division stops. [8] Hayflick's discovery later contributed to the determination of the biological roles of telomeres. [9] Hayflick claimed that the finite capacity of normal human cells to replicate was an expression of aging or senescence at the cellular level. [2] [4] [8]

During this period of research, Hayflick also discovered that if cells were properly stored in a freezer, cells would remain viable and that an enormous number of cells could be produced from a single starting culture. One of the cell strains that Hayflick isolated, which he named WI-38, was found to be free of contaminating viruses, unlike the primary monkey kidney cells then in use for virus vaccine production. [4] In addition, WI-38 cells could be frozen, then thawed and exhaustively tested. These advantages led to WI-38 quickly replacing primary monkey kidney cells for human virus vaccine production. [6] [7] [10] WI-38 has also been used for research on numerous aspects of normal human cell biology. [7] [8] [10]

Applications

WI-38 was invaluable to early researchers, especially those studying virology and immunology, since it was a readily available cell line of normal human tissue. Unlike the HeLa cell line, which were cancerous cells, WI-38 was a normal human cell population. Researchers in labs across the globe have since used WI-38 in their discoveries, most notably Hayflick in his development of human virus vaccines. [6] Infected WI-38 cells secrete the virus, and can be cultured in large volumes suitable for commercial production. [2]

Virus vaccines produced in WI-38 have prevented disease or saved the lives of billions of people. [6] [7] Vaccines produced in WI-38 include those made against adenoviruses, rubella, measles, mumps, varicella zoster, poliovirus, hepatitis A and rabies. [5] [6] [7] [10]

Genome sequence

The WI-38 cell line was one of the first cell lines whose diploid genome was sequenced. [11] This is critical because most human genome sequences have not been resolved to chromosome level, that is, it remained largely unclear which genetic variant is on which of the two chromatids. Besides being an important cell line for experimental studies (e.g. on aging), the WI-38 line is believed to have remained diploid since it was originally established in 1961. Nearly 60 years later, karyotyping by Soifer et al. (2020) showed that the WI-38 genome has not acquired major rearrangements such as translocations. More importantly, the de novo phased assembly confirms that the genome has in fact remained diploid and retained its heterozygosity throughout. It is therefore a good model for genome sequencing and serves as another reference genome. [11]

See also

Related Research Articles

<span class="mw-page-title-main">Telomere</span> Region of repetitive nucleotide sequences on chromosomes

A telomere is a region of repetitive nucleotide sequences associated with specialized proteins at the ends of linear chromosomes. Telomeres are a widespread genetic feature most commonly found in eukaryotes. In most, if not all species possessing them, they protect the terminal regions of chromosomal DNA from progressive degradation and ensure the integrity of linear chromosomes by preventing DNA repair systems from mistaking the very ends of the DNA strand for a double-strand break.

<span class="mw-page-title-main">Telomerase</span> Telomere-restoring protein active in the most rapidly dividing cells

Telomerase, also called terminal transferase, is a ribonucleoprotein that adds a species-dependent telomere repeat sequence to the 3' end of telomeres. A telomere is a region of repetitive sequences at each end of the chromosomes of most eukaryotes. Telomeres protect the end of the chromosome from DNA damage or from fusion with neighbouring chromosomes. The fruit fly Drosophila melanogaster lacks telomerase, but instead uses retrotransposons to maintain telomeres.

<span class="mw-page-title-main">HeLa</span> Oldest cultured human cell line (1951)

HeLa is an immortalized cell line used in scientific research. It is the oldest human cell line and one of the most commonly used. HeLa cells are durable and prolific, allowing for extensive applications in scientific study. The line is derived from cervical cancer cells taken on 8 February 1951, from Henrietta Lacks, a 31-year-old African American mother of five, after whom the line is named. Lacks died of cancer on 4 October 1951.

<span class="mw-page-title-main">Poliovirus</span> Enterovirus

Poliovirus, the causative agent of polio, is a serotype of the species Enterovirus C, in the family of Picornaviridae. There are three poliovirus serotypes: types 1, 2, and 3.

Mycoplasma pneumoniae is a very small bacterium in the class Mollicutes. It is a human pathogen that causes the disease mycoplasma pneumonia, a form of atypical bacterial pneumonia related to cold agglutinin disease. M. pneumoniae is characterized by the absence of a peptidoglycan cell wall and resulting resistance to many antibacterial agents. The persistence of M. pneumoniae infections even after treatment is associated with its ability to mimic host cell surface composition.

Leonard Hayflick is a Professor of Anatomy at the UCSF School of Medicine, and was Professor of Medical Microbiology at Stanford University School of Medicine. He is a past president of the Gerontological Society of America and was a founding member of the council of the National Institute on Aging (NIA). The recipient of a number of research prizes and awards, including the 1991 Sandoz Prize for Gerontological Research, he has studied the aging process for more than fifty years. He is known for discovering that normal human cells divide for a limited number of times in vitro. This is known as the Hayflick limit. His discoveries overturned a 60-year old dogma that all cultured cells are immortal. Hayflick demonstrated that normal cells have a memory and can remember at what doubling level they have reached. He demonstrated that his normal human cell strains were free from contaminating viruses. His cell strain WI-38 soon replaced primary monkey kidney cells and became the substrate for the production of most of the world's human virus vaccines. Hayflick discovered that the etiological agent of primary atypical pneumonia was not a virus as previously believed. He was the first to cultivate the causative organism called a mycoplasma, the smallest free-living organism, which Hayflick isolated on a unique culture medium that bears his name. He named the organism Mycoplasma pneumoniae.

G<sub>0</sub> phase Quiescent stage of the cell cycle in which the cell does not divide

The G0 phase describes a cellular state outside of the replicative cell cycle. Classically, cells were thought to enter G0 primarily due to environmental factors, like nutrient deprivation, that limited the resources necessary for proliferation. Thus it was thought of as a resting phase. G0 is now known to take different forms and occur for multiple reasons. For example, most adult neuronal cells, among the most metabolically active cells in the body, are fully differentiated and reside in a terminal G0 phase. Neurons reside in this state, not because of stochastic or limited nutrient supply, but as a part of their developmental program.

<span class="mw-page-title-main">Cell culture</span> Process by which cells are grown under controlled conditions

Cell culture or tissue culture is the process by which cells are grown under controlled conditions, generally outside of their natural environment. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. This technique is also called micropropagation. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator. These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (CO2, O2), and regulates the physio-chemical environment (pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as a suspension culture. This is typically facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants. The lifespan of most cells is genetically determined, but some cell-culturing cells have been “transformed” into immortal cells which will reproduce indefinitely if the optimal conditions are provided.

Biological immortality is a state in which the rate of mortality from senescence is stable or decreasing, thus decoupling it from chronological age. Various unicellular and multicellular species, including some vertebrates, achieve this state either throughout their existence or after living long enough. A biologically immortal living being can still die from means other than senescence, such as through injury, poison, disease, predation, lack of available resources, or changes to environment.

<span class="mw-page-title-main">Hayflick limit</span> Limit to divisions of a normal human cell

The Hayflick limit, or Hayflick phenomenon, is the number of times a normal somatic, differentiated human cell population will divide before cell division stops. However, this limit does not apply to stem cells.

<span class="mw-page-title-main">Wistar Institute</span> American biomedical research institute

The Wistar Institute is an independent, nonprofit research institution in biomedical science with special focuses in oncology, immunology, infectious disease and vaccine research. Located on Spruce Street in Philadelphia’s University City neighborhood, Wistar was founded in 1892 as a nonprofit institution to focus on biomedical research and training.

<span class="mw-page-title-main">Cellular senescence</span> Phenomenon characterized by the cessation of cell division

Cellular senescence is a phenomenon characterized by the cessation of cell division. In their experiments during the early 1960s, Leonard Hayflick and Paul Moorhead found that normal human fetal fibroblasts in culture reach a maximum of approximately 50 cell population doublings before becoming senescent. This process is known as "replicative senescence", or the Hayflick limit. Hayflick's discovery of mortal cells paved the path for the discovery and understanding of cellular aging molecular pathways. Cellular senescence can be initiated by a wide variety of stress inducing factors. These stress factors include both environmental and internal damaging events, abnormal cellular growth, oxidative stress, autophagy factors, among many other things.

<span class="mw-page-title-main">Rabies vaccine</span> Vaccines to prevent rabies in humans and animals

The rabies vaccine is a vaccine used to prevent rabies. There are several rabies vaccines available that are both safe and effective. Vaccinations must be administered prior to rabies virus exposure or within the latent period after exposure to prevent the disease. Transmission of rabies virus to humans typically occurs through a bite or scratch from an infectious animal, but exposure can occur through indirect contact with the saliva from an infectious individual.

<span class="mw-page-title-main">Rubella vaccine</span> Vaccine used to prevent rubella

Rubella vaccine is a vaccine used to prevent rubella. Effectiveness begins about two weeks after a single dose and around 95% of people become immune. Countries with high rates of immunization no longer see cases of rubella or congenital rubella syndrome. When there is a low level of childhood immunization in a population it is possible for rates of congenital rubella to increase as more women make it to child-bearing age without either vaccination or exposure to the disease. Therefore, it is important for more than 80% of people to be vaccinated. By introducing rubella containing vaccines, rubella has been eradicated in 81 nations, as of mid-2020.

<span class="mw-page-title-main">Robert M. Chanock</span> American pediatrician and virologist

Robert Merritt Chanock was an American pediatrician and virologist who made major contributions to the prevention and treatment of childhood respiratory infections in more than 50 years spent at the National Institute of Allergy and Infectious Diseases.

<span class="mw-page-title-main">The Hallmarks of Cancer</span> 2000 paper by Hanahan and Weinberg

The hallmarks of cancer were originally six biological capabilities acquired during the multistep development of human tumors and have since been increased to eight capabilities and two enabling capabilities. The idea was coined by Douglas Hanahan and Robert Weinberg in their paper "The Hallmarks of Cancer" published January 2000 in Cell.

Judith Campisi is an American biochemist and cell biologist. She is a professor of biogerontology at the Buck Institute for Research on Aging. She is also a member of the SENS Research Foundation Advisory Board and an adviser at the Lifeboat Foundation. She is co-editor in chief of the Aging Journal, together with Mikhail Blagosklonny and David Sinclair, and founder of the pharmaceutical company Unity Biotechnology. She is listed in Who's Who in Gerontology. She is widely known for her research on how senescent cells influence aging and cancer — in particular the Senescence Associated Secretory Phenotype (SASP).

<span class="mw-page-title-main">MRC-5</span> Cell line

MRC-5 is a diploid cell culture line composed of fibroblasts, originally developed from the lung tissue of a 14-week-old aborted Caucasian male fetus. The cell line was isolated by J.P. Jacobs and colleagues in September 1966 from the seventh population doubling of the original strain, and MRC-5 cells themselves are known to reach senescence in around 45 population doublings.

The use of fetal tissue in vaccine development is the practice of researching, developing, and producing vaccines through growing viruses in cultured (laboratory-grown) cells that were originally derived from human fetal tissue. Since the cell strains in use originate from abortions, there has been opposition to the practice and the resulting vaccines on religious and moral grounds.

Aging is characterized by a progressive loss of physiological integrity, leading to impaired function and increased vulnerability to death. The hallmarks of aging are the types of biochemical changes that occur in all organisms that experience biological aging and lead to a progressive loss of physiological integrity, impaired function and, eventually, death. They were first listed in a landmark paper in 2013 to conceptualize the essence of biological aging and its underlying mechanisms.

References

  1. "WI-38 (ATCC® CCL-75™)".
  2. 1 2 3 4 5 Hayflick L, Moorhead PS (December 1961). "The serial cultivation of human diploid cell strains". Experimental Cell Research. 25 (3): 585–621. doi:10.1016/0014-4827(61)90192-6. PMID   13905658.
  3. 1 2 Gorvett Z. "The controversial cells that saved 10 million lives". www.bbc.com. Retrieved 2020-12-09.
  4. 1 2 3 4 Hayflick L (March 1965). "The Limited in vitro Lifetime of Human Diploid Cell Strains". Experimental Cell Research. 37 (3): 614–36. doi:10.1016/0014-4827(65)90211-9. PMID   14315085.
  5. 1 2 Fletcher MA, Hessel L, Plotkin SA (1998). "Human diploid cell strains (HDCS) viral vaccines". Developments in Biological Standardization. 93: 97–107. PMID   9737384.
  6. 1 2 3 4 5 Olshansky SJ, Hayflick L (2 March 2017). "The Role of the WI-38 Cell Strain in Saving Lives and Reducing Morbidity". AIMS Public Health. 4 (2): 127–138. doi:10.3934/publichealth.2017.2.127. PMC   5689800 . PMID   29546209.
  7. 1 2 3 4 5 Wadman M (June 2013). "Medical research: cell division". Nature. 498 (7455): 422–6. Bibcode:2013Natur.498..422W. doi:10.1038/498422a. PMID   23803825.
  8. 1 2 3 Shay JW, Wright WE (October 2000). "Hayflick, his limit, and cellular ageing" (PDF). Nature Reviews. Molecular Cell Biology. 1 (1): 72–6. doi:10.1038/35036093. PMID   11413492. S2CID   6821048. Archived from the original on 2010-07-13.{{cite journal}}: CS1 maint: bot: original URL status unknown (link)
  9. Holliday R (2012). "Telomeres and telomerase: the commitment theory of cellular ageing revisited". Science Progress. 95 (Pt 2): 199–205. doi: 10.3184/003685012X13361526995348 . PMC   10365536 . PMID   22893980. S2CID   20557366.
  10. 1 2 3 Hayflick L. "Errors in the "Vaccine Race" Book" (PDF). Archived from the original (PDF) on 2019-01-22. Retrieved 2018-04-24.
  11. 1 2 Soifer L, Fong NL, Yi N, Ireland AT, Lam I, Sooknah M, et al. (September 2020). "Fully Phased Sequence of a Diploid Human Genome Determined de Novo from the DNA of a Single Individual". G3. 10 (9): 2911–2925. doi:10.1534/g3.119.400995. PMC   7466960 . PMID   32631951.