Differentiation of newborn rat adipocyte precursors in defined serum-free medium |
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Authors: | Ginette Serrero Dianne Mills |
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Institution: | (1) W. Alton Jones Cell Science Center, Inc., 10 Old Barn Road, 12946 Lake Placid, New York |
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Abstract: | Summary Newborn rat adipocyte precursors, isolated from inguinal fat pads of 2 day-old NBR rats proliferate and undergo adipose differentiation
in defined medium in the absence of serum when cultivated on polylysine coated dishes in DME-F12 medium supplemented with
fibronectin, insulin, transferrin and FGF. After 7 days in culture in these conditions, 90% of the cells have undergone differentiation
as measured by the increase of G3PDH specific activity and by the accumulation of triglycerides in their cytoplasm. In contrast,
the cells cultivated in the presence of 10% fetal bovine serum, have a limited ability to differentiate. These results indicate
that newborn rat adipocyte precursors from inguinal fat pads do not require the presence of an undefined adipogenic factor
in order to differentiate in culture. In contrast, proliferation and differentiation are dependent on the presence of insulin
in the culture medium. Moreover, the data presented in this paper show that the rat adipocyte precursor culture represents
a rapid and reproducible system for investigating the processes of adipose tissue development and for studying the negative
and positive regulators of the adipose differentiation in a controlled environment.
This work was supported by grants from the Juvenile Diabetes Foundation, File #185221 and from the National Institutes of
Health 1 PO1 CA37589.
Editor’s Statement This paper extends to primary cultures the serum-free methods previously applied to studies of adipocyte
differentiation in established lines. The observation that serum can block differentiation in this system suggests the existence
of previously unrecognized circulating plasma or platelet factors affecting adipocyte differentiation, and the model developed
provides an assay for the identification of these factors. |
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Keywords: | differentiation adipocyte precursor insulin defined medium |
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