Production of somatomedin-like activity by human adult tumor-derived, transformed, and normal cell cultures and by cultured rat hepatocytes: effects of culture conditions and of epidermal growth factor (urogastrone)

1984 ◽  
Vol 62 (12) ◽  
pp. 1343-1350 ◽  
Author(s):  
Paul R. Atkison ◽  
L. Joseph Hayden ◽  
R. Marvin Bala ◽  
Morley D. Hollenberg
Keyword(s):  
Human Skin ◽  
Cell Cultures ◽  
Culture Medium ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Kb Cells ◽  
Serum Free ◽  
Epidermal Growth ◽  
Adult Human Skin

We have measured the production of a basic-somatomedin-like activity (SLA) by a variety of human tumor-derived, transformed, and normal postnatal cell cultures; and we have compared the production of SLA by these cell types with the production of SLA by adult rat hepatocytes cultured in serum-free medium. Cells derived from a human epidermoid carcinoma (KB), a pancreatic carcinoma (Panc-1), a Simian virus 40 transformed adult human skin-derived cell line (SV40 fibroblasts), and a normal adult human skin-derived fibroblast line released SLA when cultured in a serum-free growth medium. No SLA was recovered from the culture medium of human choriocarcinoma-derived cells (BeWo) or of a human lymphoblastoid cell line (IM-9). The production of SLA by rat hepatocytes cultured in serum-free medium appeared to exceed the production of SLA by the other cell cultures. In cultures of KB cells, SV40 fibroblasts, and rat hepatocytes, the production of SLA depended on the frequency with which the growth medium was renewed; in general, the highest rates of SLA production were observed when the medium was renewed every 48–72 h. The presence of mouse epidermal growth factor (urogastrone) (EGF-URO) in the serum-free culture medium stimulated the production of SLA by KB cells and by rat hepatocytes, but did not increase SLA production by normal or by SV40-transfonned human skin-derived fibroblasts. We conclude that tumor-derived cells are capable of producing somatomedin-like activity and that the production of SLA by such cells can be subject to controls (nutrient availability, EGF-URO stimulation) that regulate SLA production, either by normal adult tissues, like liver, or by a variety of normal embryonic tissues.

Receptors for epidermal growth factor (urogastrone) and insulin in primary cultures of rat hepatocytes maintained in serum-free medium

1986 ◽  
Vol 64 (8) ◽  
pp. 803-810 ◽  
Author(s):  
M. O'Connor-McCourt ◽  
M. Soley ◽  
L. J. Hayden ◽  
M. D. Hollenberg
Keyword(s):  
Growth Factor ◽  
Molecular Mass ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Cross Link ◽  
Serum Free ◽  
Epidermal Growth

We have analyzed the receptors for epidermal growth factor (urogastrone) (EGF-URO) and insulin in primary cultures of adult rat hepatocytes maintained for up to 3 weeks on human placental cell matrix in serum-free defined medium. Cross-link labeling experiments revealed that the insulin receptor, partially damaged by the collagenase isolation procedure, was rapidly regenerated to yield an intact receptor. In contrast, cross-link labeling of the EGF-URO receptor revealed that, upon prolonged culture, there was a progressive disappearance of the high molecular mass (175 kilodaltons (kDa)) receptor form, and an appearance of low molecular mass receptor species (130 and 105 kDa). After 3 weeks of culture, the low molecular mass receptor forms accounted for all of the labeled EGF-URO receptor present in the cells. Measurements of EGF-URO binding indicated that the number of EGF-URO binding sites per cell (2.0 × 105 ± 0.3 × 105) did not change during the 3 weeks of culture. However, there was a decrease in EGF-URO binding affinity, reflected by an increase in the KD from 0.6 to 3.0 nM. At zero time and after 3 weeks in culture, Scatchard plots of the binding data were linear; at intermediate time points, the plots were curvilinear. Despite the changes in the EGF-URO receptor that occurred, cells were still responsive to EGF-URO in terms of the inhibition of acetate incorporation into lipid. The ED50 for EGF-URO (about 0.2 nM) was the same for short-term cultures (48 h) as for cells maintained in culture for 3 weeks. We conclude that the long-term culture of hepatocytes in serum-free medium yields an altered low molecular form of the EGF-URO receptor that is, nonetheless, functional. The study points to differential changes in receptors for peptide hormones that may occur in long-term hepatocyte cultures and illustrates the feasibility of using such cultures for metabolic studies of the actions of EGF-URO.

Characterization of neurons from adult human skin-derived precursors in serum-free medium : a PCR array and immunocytological analysis

2012 ◽  
Vol 21 (3) ◽  
pp. 195-200 ◽  
Author(s):  
Nicolas Lebonvallet ◽  
Nicholas Boulais ◽  
Christelle Le Gall ◽  
Jeremy Chéret ◽  
Ulysse Pereira ◽  
...  
Keyword(s):  
Human Skin ◽  
Pcr Array ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Adult Human ◽  
Adult Human Skin

Reactivity of monoclonal anti-human skin basal cell antibody to cultured keratinocytes in serum-free medium

1990 ◽  
Vol 1 (1) ◽  
pp. 47-55
Author(s):  
F FURUKAWA ◽  
D NORRIS ◽  
M KASHIHARASAWAMI ◽  
M LYONS ◽  
M UEDA ◽  
...  
Keyword(s):  
Human Skin ◽  
Basal Cell ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cultured Keratinocytes ◽  
Cell Antibody

Cell surface heparan sulphate and adhesive property of sublines of rat ascites hepatoma AH7974

1988 ◽  
Vol 90 (4) ◽  
pp. 683-689 ◽  
Author(s):  
A. Kimura ◽  
T. Kawaguchi ◽  
T. Ono ◽  
A. Sakuma ◽  
Y. Yokoya ◽  
...  
Keyword(s):  
Cell Surface ◽  
Culture Medium ◽  
Heparan Sulphate ◽  
Soluble Form ◽  
Foetal Calf Serum ◽  
Serum Free Medium ◽  
Free Medium ◽  
Ascites Hepatoma ◽  
Serum Free ◽  
Rat Ascites Hepatoma

Two variants (74AD and 74FL) established from rat ascites hepatoma AH7974 were examined for the production of glycosaminoglycans in culture. There was no difference between the adhesive (74AD) and the floating (74FL) variants in quantity of glycosaminoglycans produced by their cultivation in minimum essential medium supplemented with 10% foetal calf serum. However, they were distinctly different in the distribution patterns of heparan sulphate. In 74FL, about 70% of total heparan sulphate was found in the culture medium in soluble form, whereas in 74AD, only 7% was found in the medium and the rest was in the cell-substratum complex. In a serum-free medium, 74AD cells grew without adhering to the substratum. After cultivation, more than 90% of total heparan sulphate was found in the cell-associated fractions and the rest in the substratum fractions. No heparan sulphate was detected in the culture medium. On the other hand, 74FL cells released heparan sulphate to the serum-free medium as much as to the serum-containing medium. The increase in amount of heparan sulphate in the culture medium of 74FL cells was supposed to be caused by failure of the cells to deposit heparan sulphate at the cell surface and not caused by increased production. Cell-substratum adhesion mechanisms involving cell surface heparan sulphate (heparan sulphate proteoglycan) and some serum intermediate(s) are discussed for 74AD cells.

scholarly journals Stimulierung von Kulturen embryonaler Rattenzellen durch eine Proteinfraktion aus fötalem Kälberserum / Stimulation of Embryonic Rat Cells in Culture by a Protein Fraction Isolated from Fetal Calf Serum

1971 ◽  
Vol 26 (10) ◽  
pp. 1045-1048 ◽  
Author(s):  
Dieter F. Hülser ◽  
Werner Frank
Keyword(s):  
Cell Cycle ◽  
Culture Medium ◽  
Fetal Calf Serum ◽  
Surface Membrane ◽  
Calf Serum ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Membrane Potential Difference ◽  
Stimulation Of

Normal embryonic rat cells incubated in serum-free medium accumulate in G1-phase of the cell cycle. On addition of a growth-stimulating protein isolated from fetal calf serum they are triggered to proceed through the cycle, and they resume DNA-synthesis 15 to 20 hours later. In this paper it is demonstrated that the surface membrane potential difference (PD) decreases immediately after changing serum-free medium against culture medium containing either calf serum or the isolated serum protein; the original PD is restored 2 to 3 hours later. Serumprotein without growthstimulating activity does not affect the PD.A permanent rat cell line which grows independently of serum also has been tested. The PD of these cells is not significantly influenced by calf serum.

Long-term culture of rat hepatocytes on porous membranes in hormonally defined serum-free medium

1993 ◽  
Vol 7 (4) ◽  
pp. 453-459 ◽  
Author(s):  
C. Guery ◽  
J.P. Stepniewski ◽  
B. Vannier ◽  
R. Fournex ◽  
G. Lorenzon
Keyword(s):  
Rat Hepatocytes ◽  
Porous Membranes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Long Term Culture
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