A Chemically Defined Medium for Growth of Animal Cells in Suspension

Abstract

identical in the 2 specimens indicating the absence of a selective proliferation of nonlabeled cells.In conclusion it appears that liver regeneration in contradistinction to red cell regeneration is the result of multiplication of the general liver cell population rather than multiple divisions of a small segment of that population. Summary. A study of hepatic regenerationin rats was carried out to see whether the stimulus of partial hepatectomy promotes proliferation of parenchymal liver cells or, in analogy with red cell regeneration, promotes the differentiation of stem cells to new hepatic parenchymal cells. By labeling a large proportion of hepatic cells with tritiated thymidine it was shown that the labeled cells rather than the presumably unlabeled stem cells were responsible for hepatic regeneration following partial hepatectomy.The growth of tissue cells (L cells) in suspension in a chemically defined medium, NCTC 109 with methylcellulose, has been reported by Bryant et aZ.(l). Bakken et aZ. ( 2 ) reported that this medium also supported the growth of suspension cultures of a line of human skin cells. Merchant and Hellman ( 3 ) have recently described the growth of L-M strain mouse cells in suspension culture using chemically defined 2X Eagle's medium. A serum-free medium containing lactalbumin hydrolyzate described by Higuchi (4) that supports the growth of several cell lines in monolayer cultures was modified for the growth of tissue cells in suspension culture by Tribble and Higuchi( 5 ) .This report presents a chemically defined medium formulated as a result of experiments with a cat kidney cell line, and its successful application for the growth of 5 other cell lines. Materials and methods. Preparation of the medium.Salts (except sodium bicarbonate), glucose, amino acids, and glutamine were dissolved together in hot distilled water. Vitamins, stored at -20°C as a lOOX stock solution, were added to the cooled mixture. Sodium bicarbonate and phenol red dissolved together in a small amount of water were added and the resulting solution was sterilized by filtration through a 0.2 p membrane filter and stored at 5°C as a 5X solution. Prior to inoculation, insulin, methylcellulose, streptomycin and penicillin were added aseptically and the final volume adjusted by addition of sterile distilled water. Methylcellulose was stored as a sterile 2% solution; antibiotics were stored at -20°C at lOOX concentrations. The medium used for determining amino acid requirements was prepared by adding aseptically to the filtered salts-vita,min solution individual amino acids ( 1 W X by guest on July 27, 2015 ebm.sagepub.com Downloaded from

Systems approach to investigating host-pathogen interactions in infections with the biothreat agent Francisella. Constraints-based model of Francisella tularensis

“…F. tularensis was also able to grow in Chamberlain media on amino acids without an additional carbohydrate source. Increasing the amino acid concentration increased the growth rates as has been reported [40,41]. However, the specific growth rates were slower and the final biomass concentration was much lower (Figure 4B).…”

The complete genome sequence of Francisella tularensis, the causative agent of tularemia

“…18) and was also reported to support growth of SCHU S4 (ref. 19). F. tularensis strain SCHU S4 also has a requirement for cysteine 20 , which seems to be due to a nonfunctional pathway for sulfate assimilation resulting from a pseudogene (missing start codon) encoding adenylylsulfate kinase (EC 2.7.1.25).…”

Utilization and formation of amino acids by chicken epiphyseal chondrocytes: Comparative studies with cultured cells and native cartilage tissue

“…Growth of mammalian cells in chemically defined synthetic culture media have been reported by numerous investigators (Eagle, 1959;Dulbecco and Freeman, 1959;Biggers et al, 1961;Ham, 1965;Moore et al, 1967;Nagle, 1968;Yamane et al, 1968;McKeehan et al, 1976;Burks and Peck, 1978). However, knowledge of the specific amino acid requirements of individual cell types in culture, and of the relationship between these and the physiological environment around cells in vivo (Munro and Portugal, 1972) is lacking.…”