Immortalised cell line

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Immortalised cell line
HeLa-IV.jpg
Scanning electron micrograph of an apoptotic HeLa cell. Zeiss Merlin HR-SEM.
HeLa Hoechst 33258.jpg
HeLa cells, an example of an immortalised cell line. DIC image, DNA stained with Hoechst 33258.
Anatomical terminology

An immortalised cell line is a population of cells from a multicellular organism that would normally not proliferate indefinitely but, due to mutation, have evaded normal cellular senescence and instead can keep undergoing division. The cells can therefore be grown for prolonged periods in vitro . The mutations required for immortality can occur naturally or be intentionally induced for experimental purposes. Immortal cell lines are a very important tool for research into the biochemistry and cell biology of multicellular organisms. Immortalised cell lines have also found uses in biotechnology.

Contents

An immortalised cell line should not be confused with stem cells, which can also divide indefinitely, but form a normal part of the development of a multicellular organism.

Relation to natural biology and pathology

There are various immortal cell lines. Some of them are normal cell lines (e.g. derived from stem cells). Other immortalised cell lines are the in vitro equivalent of cancerous cells. Cancer occurs when a somatic cell that normally cannot divide undergoes mutations that cause deregulation of the normal cell cycle controls, leading to uncontrolled proliferation. Immortalised cell lines have undergone similar mutations, allowing a cell type that would normally not be able to divide to be proliferated in vitro. The origins of some immortal cell lines – for example, HeLa human cells – are from naturally occurring cancers. HeLa, the first immortal human cell line on record to be successfully isolated and proliferated by a laboratory, was taken from Henrietta Lacks in 1951 at Johns Hopkins Hospital in Baltimore, Maryland. [1]

Role and uses

Immortalised cell lines are widely used as a simple model for more complex biological systems – for example, for the analysis of the biochemistry and cell biology of mammalian (including human) cells. [2] The main advantage of using an immortal cell line for research is its immortality; the cells can be grown indefinitely in culture. This simplifies analysis of the biology of cells that may otherwise have a limited lifetime.[ citation needed ]

Immortalised cell lines can also be cloned, giving rise to a clonal population that can, in turn, be propagated indefinitely. This allows an analysis to be repeated many times on genetically identical cells, which is desirable for repeatable scientific experiments. The alternative, performing an analysis on primary cells from multiple tissue donors, does not have this advantage.[ citation needed ]

Immortalised cell lines find use in biotechnology, where they are a cost-effective way of growing cells similar to those found in a multicellular organism in vitro. The cells are used for a wide variety of purposes, from testing toxicity of compounds or drugs to production of eukaryotic proteins.[ citation needed ]

Limitations

Changes from nonimmortal origins

While immortalised cell lines often originate from a well-known tissue type, they have undergone significant mutations to become immortal. This can alter the biology of the cell and must be taken into consideration in any analysis. Further, cell lines can change genetically over multiple passages, leading to phenotypic differences among isolates and potentially different experimental results depending on when and with what strain isolate an experiment is conducted. [3]

Contamination with other cells

Many cell lines that are widely used for biomedical research have been contaminated and overgrown by other, more aggressive cells. For example, supposed thyroid lines were actually melanoma cells, supposed prostate tissue was actually bladder cancer, and supposed normal uterine cultures were actually breast cancer. [4]

Methods of generation

There are several methods for generating immortalised cell lines: [5]

  1. Isolation from a naturally occurring cancer. This is the original method for generating an immortalised cell line. A major example is human HeLa, a line derived from cervical cancer cells taken on February 8, 1951 from Henrietta Lacks, a 31-year-old African-American mother of five, who died of cancer on October 4, 1951. [6]
  2. Introduction of a viral gene that partially deregulates the cell cycle (e.g., the adenovirus type 5 E1 gene was used to immortalise the HEK 293 cell line; the Epstein–Barr virus can immortalise B lymphocytes by infection [7] ).
  3. Artificial expression of key proteins required for immortality, for example telomerase which prevents degradation of chromosome ends during DNA replication in eukaryotes. [8]
  4. Hybridoma technology, specifically used for generating immortalised antibody-producing B cell lines, where an antibody-producing B cell is fused with a myeloma (B cell cancer) cell. [9]

Examples

There are several examples of immortalised cell lines, each with different properties. Most immortalised cell lines are classified by the cell type they originated from or are most similar to biologically

See also

Related Research Articles

In cellular biology, the term somatic is derived from the French somatique which comes from Ancient Greek σωματικός, and σῶμα is often used to refer to the cells of the body, in contrast to the reproductive (germline) cells, which usually give rise to the egg or sperm. These somatic cells are diploid, containing two copies of each chromosome, whereas germ cells are haploid, as they only contain one copy of each chromosome. Although under normal circumstances all somatic cells in an organism contain identical DNA, they develop a variety of tissue-specific characteristics. This process is called differentiation, through epigenetic and regulatory alterations. The grouping of similar cells and tissues creates the foundation for organs.

<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">Henrietta Lacks</span> African-American woman (1920–1951), source of HeLa immortal cell line

Henrietta Lacks was an African-American woman whose cancer cells are the source of the HeLa cell line, the first immortalized human cell line and one of the most important cell lines in medical research. An immortalized cell line reproduces indefinitely under specific conditions, and the HeLa cell line continues to be a source of invaluable medical data to the present day.

<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 February 8, 1951, from Henrietta Lacks, a 31-year-old African American mother of five, after whom the line is named. Lacks died of cancer on October 4, 1951.

<span class="mw-page-title-main">Germline</span> Population of a multicellular organisms cells that pass on their genetic material to the progeny

In biology and genetics, the germline is the population of a multicellular organism's cells that develop into germ cells. In other words, they are the cells that form gametes, which can come together to form a zygote. They differentiate in the gonads from primordial germ cells into gametogonia, which develop into gametocytes, which develop into the final gametes. This process is known as gametogenesis.

<span class="mw-page-title-main">Tissue culture</span> Growth of tissues or cells in an artificial medium separate from the parent organism

Tissue culture is the growth of tissues or cells in an artificial medium separate from the parent organism. This technique is also called micropropagation. 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 term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. This is possible only in certain conditions. It also requires more attention. It can be done only in genetic labs with various chemicals.

<span class="mw-page-title-main">Hep G2</span>

Hep G2 is a human liver cancer cell line.

<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. After 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.

A mitogen is a small bioactive protein or peptide that induces a cell to begin cell division, or enhances the rate of division (mitosis). Mitogenesis is the induction (triggering) of mitosis, typically via a mitogen.

Carcinogenesis, also called oncogenesis or tumorigenesis, is the formation of a cancer, whereby normal cells are transformed into cancer cells. The process is characterized by changes at the cellular, genetic, and epigenetic levels and abnormal cell division. Cell division is a physiological process that occurs in almost all tissues and under a variety of circumstances. Normally, the balance between proliferation and programmed cell death, in the form of apoptosis, is maintained to ensure the integrity of tissues and organs. According to the prevailing accepted theory of carcinogenesis, the somatic mutation theory, mutations in DNA and epimutations that lead to cancer disrupt these orderly processes by interfering with the programming regulating the processes, upsetting the normal balance between proliferation and cell death. This results in uncontrolled cell division and the evolution of those cells by natural selection in the body. Only certain mutations lead to cancer whereas the majority of mutations do not.

<span class="mw-page-title-main">Stem-cell line</span> Culture of stem cells that can be propagated indefinitely

A stem cell line is a group of stem cells that is cultured in vitro and can be propagated indefinitely. Stem cell lines are derived from either animal or human tissues and come from one of three sources: embryonic stem cells, adult stem cells, or induced pluripotent stem cells. They are commonly used in research and regenerative medicine.

<span class="mw-page-title-main">George Otto Gey</span> American cell biologist (1899–1970)

George Otto Gey was the cell biologist at Johns Hopkins Hospital who is credited with propagating the HeLa cell line from Henrietta Lacks' cervical tumor. He spent over 35 years developing numerous scientific breakthroughs under the Johns Hopkins Medical School and Hospital.

<span class="mw-page-title-main">Clone (cell biology)</span> Group of identical cells that share a common ancestry

A clone is a group of identical cells that share a common ancestry, meaning they are derived from the same cell.

HaCaT is a spontaneously transformed aneuploid immortal keratinocyte cell line from adult human skin, widely used in scientific research. HaCaT cells are utilized for their high capacity to differentiate and proliferate in vitro. Their use in research allows for the characterization of human keratinocyte using a model that is reproducible and addresses issues such as short culture lifespan and variations between cell lines that would otherwise be encountered. These cells have allowed the characterization of several processes, such as their utilization as a model system for vitamin D3 metabolism in the skin.

HT-29 is a human colon cancer cell line used extensively in biological and cancer research.

A somatic mutation is a change in the DNA sequence of a somatic cell of a multicellular organism with dedicated reproductive cells; that is, any mutation that occurs in a cell other than a gamete, germ cell, or gametocyte. Unlike germline mutations, which can be passed on to the descendants of an organism, somatic mutations are not usually transmitted to descendants. This distinction is blurred in plants, which lack a dedicated germline, and in those animals that can reproduce asexually through mechanisms such as budding, as in members of the cnidarian genus Hydra.

<span class="mw-page-title-main">Primary cell culture</span> The culture of cells from fresh tissues

Primary cell culture is the ex vivo culture of cells freshly obtained from a multicellular organism, as opposed to the culture of immortalized cell lines. In general, primary cell cultures are considered more representative of in vivo tissues than cell lines, and this is recognized legally in some countries such as the UK. However, primary cells require adequate substrate and nutrient conditions to thrive and after a certain number of divisions they acquire a senescent phenotype, leading to irreversible cell cycle arrest. The generation of cell lines stems from these two reasons. Primary cells can become immortalized either spontaneously or by genetic modification, at which point they become cell lines which can be subcultured indefinitely.

References

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  7. Henle W, Henle G (1980). "Epidemiologic aspects of Epstein-Barr virus (EBV)-associated diseases". Annals of the New York Academy of Sciences. 354: 326–31. doi:10.1111/j.1749-6632.1980.tb27975.x. PMID   6261650. S2CID   30025994.
  8. Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu CP, Morin GB, et al. (January 1998). "Extension of life-span by introduction of telomerase into normal human cells". Science. 279 (5349): 349–52. Bibcode:1998Sci...279..349B. doi:10.1126/science.279.5349.349. PMID   9454332.
  9. Kwakkenbos MJ, van Helden PM, Beaumont T, Spits H (March 2016). "Stable long-term cultures of self-renewing B cells and their applications". Immunological Reviews. 270 (1): 65–77. doi: 10.1111/imr.12395 . PMC   4755196 . PMID   26864105.