Abstract
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.
Full Text
The Full Text of this article is available as a PDF (1.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BRYANT J. C., EARLE W. R., PEPPERS E. V. The effect of ultracentrifugation and hyaluronidase on the filtrability of chick-embryo extract for tissue culture. J Natl Cancer Inst. 1953 Aug;14(1):189–225. [PubMed] [Google Scholar]
- Briggs R., King T. J. Transplantation of Living Nuclei From Blastula Cells into Enucleated Frogs' Eggs. Proc Natl Acad Sci U S A. 1952 May;38(5):455–463. doi: 10.1073/pnas.38.5.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CHANG R. S. M. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc Soc Exp Biol Med. 1954 Nov;87(2):440–443. doi: 10.3181/00379727-87-21406. [DOI] [PubMed] [Google Scholar]
- CIECIURA S. J., MARCUS P. I., PUCK T. T. Clonal growth in vitro of epithelial cells from normal human tissues. J Exp Med. 1956 Oct 1;104(4):615–628. doi: 10.1084/jem.104.4.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- EARLE W. R., BRYANT J. C., SCHILLING E. L. Certain factors limiting the size of the tissue culture and the development of massive cultures. Ann N Y Acad Sci. 1954 Nov 17;58(7):1000–1011. doi: 10.1111/j.1749-6632.1954.tb45887.x. [DOI] [PubMed] [Google Scholar]
- Fisher H. W., Puck T. T. ON THE FUNCTIONS OF X-IRRADIATED "FEEDER" CELLS IN SUPPORTING GROWTH OF SINGLE MAMMALIAN CELLS. Proc Natl Acad Sci U S A. 1956 Dec;42(12):900–906. doi: 10.1073/pnas.42.12.900. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Puck T. T., Fisher H. W. GENETICS OF SOMATIC MAMMALIAN CELLS : I. DEMONSTRATION OF THE EXISTENCE OF MUTANTS WITH DIFFERENT GROWTH REQUIREMENTS IN A HUMAN CANCER CELL STRAIN (HELA). J Exp Med. 1956 Sep 1;104(3):427–434. doi: 10.1084/jem.104.3.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Puck T. T., Marcus P. I. A RAPID METHOD FOR VIABLE CELL TITRATION AND CLONE PRODUCTION WITH HELA CELLS IN TISSUE CULTURE: THE USE OF X-IRRADIATED CELLS TO SUPPLY CONDITIONING FACTORS. Proc Natl Acad Sci U S A. 1955 Jul 15;41(7):432–437. doi: 10.1073/pnas.41.7.432. [DOI] [PMC free article] [PubMed] [Google Scholar]