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How do we maintain cells alive out of an organism?

Updated: Mar 10, 2020

As cell biologist, my work involves growing several cellular models of different tissue origin. Maintaining cell or tissues alive, outside of their host organism, has been and it is still representing one of the major challenges for cell biologists. Being animal and plant-derived, all cells need a precise set of nutrients to survive. In living organisms, these nutrients come from the daily intake, but isolated cells lack of the capacity of providing food for themselves. Hence, researchers have to come up with feeding strategies and schedules.


The cocktail of nutrients we administrate to the cell models are usually called “Media”. In the end of the 19th century, scientists as culinary chefs developed and improved the recipes of the cells “meals” that we currently use nowadays.


Although as different nationality signature dishes, the recipes could vary in the amount or the presence of particular ingredients, scientists agreed that the main diet of the cell stringently has to contain:

· Salts, to maintain the physiological balance of fluids within and outside the cell

· Water, as our cells are composed by 70% of water

· Sugar for energy production

· Amminoacids for protein synthesis

· Vitamins for cell functions regulations

· Hormones and growth factors to maintain cell proliferation




Although the recipe seems simple, the discovery of the perfect list involved many experiments scattered though time and place. Here is a summed-up chronology of the main discoveries that contributed to make long and sustained cell culture in vitro:

  1. At the end of the 19th century, Sydney Ringer figured out the formula of a solution -then named after him (the Ringer’s solution)- containing a mix of inorganic salts dissolved in water, in ratio comparable to the one of animal body fluids. Ringer successfully managed to keep a frog’s heart beating in vitro, after perfusing the heart with his solution;

  2. In 1907, however, Ross G. Harrison used lymph sacs of an adult frog to grow frog nerve fibres of a frog;

  3. At the beginning of the 20th century, the French Alexis Carrel –Nobel Prize in Physiology and Medicine in 1912 – developed a prototype of the cell culture flask for aseptic manipulation of vascular suture and transplantation of blood vessels and organs;

  4. In 1909, T. Burrows -working under Harrison’s supervision- discovered that he could use chicken blood plasma to keep chicken embryonic cells alive, instead of lymph;

  5. In 1912, Carrel achieved long-term cultivation of the chick connective tissues cells via a regular replacement of the medium;

  6. In 1911, Lewis and Warren H. Lewis demonstrated that the glucose is necessary to effective for chick embryo cell cultivation;

  7. In 1948, Albert Fischer discovered that the amminoacids components of the dialyzed blood plasma is essential to sustain cell growth in culture;

  8. In1955, Harry Eagle identified the the minimum 13 amino acids and eight vitamins are necessary for cell culture; he then developed what is known as the minimum essential medium (MEM);

  9. Later on, MEM was modified according to cell types and cultures conditions by several researchers as Renato Dulbecco and Marguerite Vogt, Clifford P. Stanners et al., and Norman N. Iscove and Fritz Melchers;

  10. In 1958, Thomas A. McCoy et al report the importance of pyruvate -essential element for cell energy production - for carcinosarcoma cells require.

  11. The medium developed by McCoy got then modified at the Roswell Park Memorial Institute (RPMI) for calcium and magnesium concentrations, and it was noted lymphocyte cultivation (RPMI 1640).

  12. As blood plasma was necessary but also expensive, it was then developed the Ham’s F-12, a completely synthetic medium of where albumin and fetuin where replaced by linoleic acid and putrescine.


REFERENCE:

  1. Tatsuma Yao and Yuta Asayama, Animal‐cell culture media: History, characteristics, and current issues, Reprod Med Biol. 2017 Apr; 16(2): 99–117. Published online 2017 Mar 21. doi: 10.1002/rmb2.12024, PMCID: PMC5661806 PMID: 29259457

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