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H W FISHER

Publications and source records attributed to H W FISHER.

15 recordsLinked to original sources

Molecular growth requirements of single mammalian cells. III. Quantitative colonial growth of single S3 cells in a medium containing synthetic small molecular constituents and two purified protein fractions.

Two purified serum protein fractions, fetuin and serum albumin, will replace whole or dialyzed serum in supporting the growth of single S3 HeLa cells in an otherwise chemically defined nutrient solution. In the serum-free medium, single S3 cells will form macroscopic colonies with essentially 100 per cent efficiency. The generation time of S3 cells in the serum-free medium is approximately 50 per cent greater than that observed in an optimal, serum-containing medium. All components of the serum-free medium are available commercially, except fetuin, which can easily be prepared in substantial quantities. The problem of the purity of the protein preparations and of their possible roles in promoting cell growth is discussed.

Animals↗

Clonal growth in vitro of human cells with fibroblastic morphology; comparison of growth and genetic characteristics of single epithelioid and fibroblast-like cells from a variety of human organs.

A methodology has been described for reliable cultivation in vitro of dispersed fibroblastic cells obtained from normal human organs. The procedure has permitted establishment of stable cell lines from almost every sample taken, among which the following organs were represented: skin, spleen, amnion, lung, liver, bone marrow, brain, muscle, and heart. Equally good growth has been achieved with cells from embryonic or adult tissues. The methods previously developed whereby single cells plated in Petri dishes grow into isolated macroscopic colonies can successfully be applied to the plating of human fibroblastic stocks. Plating efficiencies in the neighborhood of 50 to 60 per cent are readily achieved with such strains. The resulting colonies can be picked and clonal stocks established. Fibroblastic morphology is maintained in the colonies arising from every single cell of such clonal stocks. All of the single cells from epithelioid clonal strains also maintain their integrity throughout repeated subculture. Since the difference between clonal stocks of these two types is always maintained whenever the respective single cells are plated in the same medium, regardless of the previous history of these stocks, it may be concluded that a true genetic difference exists in these cell lines. In addition to the morphological differences between epithelioid and fibroblastic cell strains, the latter have more demanding nutritional requirements for single cell growth. Thus, single cells of fibroblastic lines almost never produce colonies with high efficiency unless the growth medium which is sufficient for epithelioid cells is supplemented with embryo extract, or a cell feeder layer. Fibroblastic cells are also more resistant to tryptic digestion of the bond uniting the cells to glass surfaces. By use of differential media, growth of both fibroblastic and epithelioid cells, respectively, has been obtained, from dispersed single cells obtained by trypsinization of a specimen of human embryonic lung.

Adult↗