Proliferation of 3T3-L1 preadipocytes in a completely defined serum-free medium

Abstract

We have developed a completely defined serum-free medium that supports the growth of Swiss 3T3-L1 fibroblasts to nearly the same extent as Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum. With ASF301 medium [former name, RITC 80-7; Yamane et al. Exp. Cell Res. 134, 470 (1981)], most of the 3T3-L1 cells survived for at least 10 days, but did not grow. ASF301 medium contains insulin and mouse-epidermal growth factor as growth factors, which are termed "progression factors". So we examined the effects of "competent factors" on the proliferation of 3T3-L1 preadipocytes. Growth in the medium supplemented with competent factors reached a confluent monolayer in 6-7 days. Furthermore, it was confirmed that 3T3-L1 cells grown in the serum-free medium retained the properties of differentiation into adipocytes. Our serum-free medium should be a useful tool for research on the growth and differentiation of 3T3-L1 preadipocytes.

Optimization of the differentiation of human preadipocytes in vitro

“…However, this combination of sera cannot be considered a solution to the presented immunological problems because FCS is still present. A serum‐free culture medium is certainly a promising pathway (Kawada et al, 1990; Tezel and Del Priore, 1998; Gstraunthaler, 2003) and would solve the problem of bovine contamination. However, in many cases, the serum‐free system is still not as potent as serum‐based media (Ethier, 1986; Koller et al, 1998) or additional growth factors are necessary to obtain an extend of proliferation comparable to serum‐supplementation (Kawada et al, 1990).…”

“…A serum‐free culture medium is certainly a promising pathway (Kawada et al, 1990; Tezel and Del Priore, 1998; Gstraunthaler, 2003) and would solve the problem of bovine contamination. However, in many cases, the serum‐free system is still not as potent as serum‐based media (Ethier, 1986; Koller et al, 1998) or additional growth factors are necessary to obtain an extend of proliferation comparable to serum‐supplementation (Kawada et al, 1990). Serum replacements like artificial serum containing animal‐derived proteins (Ultroser G) or vegetable‐derived proteins (Prolifix S6) to permit cellular growth and differentiation (Anselme et al, 2002) are problematic because these substances can also be a source of immune reactions and therefore cause the rejection of a transplant.…”

Carotenoids and retinoids as suppressors on adipocyte differentiation via nuclear receptors

“…3T3-L1 cells were routinely grown as described before [5]. Under these conditions, confluence was reached within 3 days.…”

The Potential for Fat Transplantation

“…More studies have been done on rat preadipocytes in recent years. Various agents have been known to induce preadipocyte differentiation in rats 25–28 . However, the clinical significance of these findings is still unknown at the present time.…”