Wednesday, July 24, 2013

A short history of cell culture media and the use of insulin

Today, we feature this article from our partners at BPI Magaine.

The ancestors of cell culture media were the balanced salt solutions (BSS) devised by early workers interested in studying isolated organs ex vivo.

Such solutions provided irrigation and supply of water, bulk inorganic anions essential for normal cell metabolism, osmotic balance (isotonicity), a carbohydrate such as glucose as energy source, and a buffering system to maintain the medium within the physiological pH range (7.2 - 7.6) usually monitored by the addition of phenol red (1).

The first BSS was composed in 1885 by Sydney Ringer (1836-1910), a British clinician and pharmacologist from University College London wh
o worked on the isolated frog heart (2).

A version containing lactate is still used for intravenous perfusion. Ringer's solution was modified by Maurice V. Tyrode (1878-1930), an American physiologist born in Besançon to study isolated kidneys. Another popular balanced salt solution was devised by John H. Hanks (1906-1990), an American microbiologist, in order to attempt to cultivate the leprosy bacillusin vitro (he never succeeded) (3). Renato Dulbecco, an Italian born (1914) American virologist (who won the Nobel Prize in Physiology and Medicine in 1975 with Howard Temin and David Baltimore for their discovery of reverse transcriptase), described the much used Phosphate Buffered Saline (PBS) (4).

However it became clear that in order to get cells to actually proliferate in vitro, additional nutrients had to be added to culture media.

The first successful attempt at tissue culture is attributed to Ross Harrison (1870-1959) at Johns Hopkins University, who explanted in 1907 tadpole tissues in clotted frog lymph as nutritional and growth support, using a hanging drop on a glass slide. He was successful in observing neurite expansions (5,6).

Building on Harrison's success, the major early figure in tissue culture was Alexis Carrel (1873-1944), a French surgeon. He moved to the University of Chicago in 1903 and obtained the Nobel Prize in Physiology and Medicine in 1912 for inventing a method to suture blood vessels. In 1906 he moved to the Rockefeller Institute for Medical Reseach and started his work on tissue culture, in which he did pioneering work (124 articles, 7,8) and dominated the field, for better or for worse, until his death. He adopted Harrison's hanging drop technique but replaced the frog lymph by a plasma clot and fed the explants serum, a salt solution and chick embryo extract, which he called "plasmatic medium", which became a standard until the 1950's. His most famous experiment was the maintenance for over 34 years  of an embryonic chicken heart explant, finally discarded two years after Carrel's death. This nurtured Carrel's firm belief that primary cells in culture were immortal given the appropriate nutrition. Definitive evidence that this is wrong was provided by Leonard Hayflick (born 1928), an American gerontologist and professor of anatomy at UCSF, who demonstrated in 1965 that normal human diploid fibroblasts do not survive in vitro for more than approximately 50 divisions ("Hayflick's limit") (9).  The survival of Carrel's culture was likely due to the daily feeding of new embryonic cells from the embryonic extract.

The first continuous culture of a rodent cell line (L929) from a single cell was generated by Wilton R. Earle at the National Cancer Institute in 1943 (10). In 1951, George O. Gey (1899 -1970) at Johns Hopkins University developed the first continuous human cancer cell line from Henrietta Lacks's cervical carcinoma, the celebrated HeLa cell line whose eagerness to grow has resulted in crosscontamination of many cell cultures…

To read the rest of the white paper, visit BPI Magaine.

Follow the jump to see the References.

1. Sigma Life Sciences Cell Culture Manual 2011-2014, p.116.
2. Miller DJ. Sydney Ringer; physiological saline, calcium and the contraction of the heart. J 
Physiol555:585-587 (2004).
3. Hanks JH. Hanks' balanced salt solution and pH control. Tissue Culture Association Manual,3,3 
4. Dulbecco R and Vogt M. Plaque formation and isolation of pure lines with polyomyelitis viruses. J 
Exp Med106:167-169 (1954).
5.Harrison R. Observation on the living developing nerve fiber. Anat Rec 1:116-128 (1907).
6.Harrison R. The outgrowth of the nerve fiber as a mode of protoplasmic movement. J Exp Zool9:787-
846 (1910).
7. Carrel A. On the permanent life of tissues outside of the organism. J Exp Med15:516-528 (1912).
8. Carrel A andLindbergh CA, The culture of whole organs. Science 81:621-623 (1935).
9. Hayflick L. The limitedin vitrolifetime of human diploid cell strains. Exp Cell Res 37:614-636 

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