Foetal Calf Serum-free media and serum alternatives

C-6 glioma cells, grown in medium supplemented with 5% delipidated foetal calf serum, were induced to enter a quiescent state by removing serum from the medium. Within 24h there was a 75–80% decline in the rate of incorporation of [14C]acetate or 3H2O into digitonin-precipitable sterols. Experiments with [3H]mevalonolactone as a labelled sterol precursor suggested that the decline in sterol synthesis was regulated primarily at a point in the pathway before the formation of mevalonate. The specific activities of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase decreased sharply in conjunction with the decline in sterol synthesis in the serum-free cultures; however, the activity of acetoacetyl-CoA thiolase was altered only slightly. The magnitude of the initial decline in reductase activity was not affected when 50-mm-NaF was included in the preincubation and assay buffers to prevent activation of physiologically inactive enzyme. However, after 6h of serum deprivation the decline in 3-hydroxy-3-methylglutaryl-CoA reductase activity was due to a decrease in the amount of latent activity. The sterol concentration in C-6 cells was unchanged after 24h in serum-free medium, although a 20% decrease in the sterol/fatty acid molar ratio occurred as a result of a small increase in the fatty-acid concentration. Incorporation of 3H2O into fatty acids was inhibited in the serum-deprived glial cells; however, this inhibition developed more slowly and was not as pronounced as the diminution in sterol synthesis. The results suggest that in C-6 glia, which resemble the glial stem cells of the developing brain, the decreased demand for membrane sterols in the quiescent state results in a decline in sterol synthesis, mediated primarily through co-ordinate changes in the activities of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase.

Download Full-text

Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100082418 ◽

1978 ◽

Vol 36 (2) ◽

pp. 310-311

Author(s):

W. Liebrich

Keyword(s):

Hela Cells ◽

Calf Serum ◽

Glass Tube ◽

Serum Free Medium ◽

Nacl Solution ◽

Free Medium ◽

Minimum Essential Medium ◽

Serum Free ◽

All Solutions ◽

Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

Download Full-text

Iron uptake and regulation of ferritin synthesis by hepatoma cells in hormone-supplemented serum-free media.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)32565-1 ◽

1983 ◽

Vol 258 (8) ◽

pp. 5248-5255

Author(s):

Y Goto ◽

M Paterson ◽

I Listowsky

Keyword(s):

Iron Uptake ◽

Hepatoma Cells ◽

Serum Free ◽

Ferritin Synthesis ◽

Free Media

Download Full-text

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Scientific Reports ◽

10.1038/s41598-021-83088-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Samatha Bhat ◽

Pachaiyappan Viswanathan ◽

Shashank Chandanala ◽

S. Jyothi Prasanna ◽

Raviraja N. Seetharam

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Population Doubling ◽

Seeding Density ◽

Population Doubling Time ◽

Differentiation Potential ◽

Serum Free ◽

Safety Issues ◽

Free Media

AbstractBone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. Although therapeutic value of MSCs is now being established through many clinical trials, issues have been raised regarding their expansion as per regulatory guidelines. Fetal bovine serum usage in cell therapy poses difficulties due to its less-defined, highly variable composition and safety issues. Hence, there is a need for transition from serum-based to serum-free media (SFM). Since SFM are cell type-specific, a precise analysis of the properties of MSCs cultured in SFM is required to determine the most suitable one. Six different commercially available low serum/SFM with two different seeding densities were evaluated to explore their ability to support the growth and expansion of BM-MSCs and assess the characteristics of BM-MSCs cultured in these media. Except for one of the SFM, all other media tested supported the growth of BM-MSCs at a low seeding density. No significant differences were observed in the expression of MSC specific markers among the various media tested. In contrary, the population doubling time, cell yield, potency, colony-forming ability, differentiation potential, and immunosuppressive properties of MSCs varied with one another. We show that SFM tested supports the growth and expansion of BM-MSCs even at low seeding density and may serve as possible replacement for animal-derived serum.

Download Full-text