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Biotechnology and Applied Biochemistry (2002) 36, (205–212) (Printed in Great Britain)

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Effects of bovine colostral ultrafiltrates on growth and differentiation of 3T3-L1 preadipocytes

Seong-Ho Lee and Kim L. Hossner¹

Department of Animal Sciences, Colorado State University, Fort Collins, CO 80523-1171, U.S.A.

Key words: colostrum, insulin-like growth factor (IGF), peroxisome-proliferator-activated receptor g (PPARg), platelet-derived growth factor (PDGF), ultrafiltration.

Abbreviations used: BCS, bovine calf serum; COL, defatted colostrum; DEX, dexamethasone; HMW, high-molecular-mass ultrafiltrate; MIX, isobutylmethylxanthine; IGF, insulin-like growth factor; MMCO, molecular-mass cut-off; LMW, low-molecular-mass ultrafiltrate; PDGF, platelet-derived growth factor; PPARg, peroxisome-proliferator-activated receptor g; TGFb, transforming growth factor-b; DMEM, Dulbecco's modified Eagle's medium; DM, differentiation medium; FBS, fetal bovine serum.

¹To whom correspondence should be addressed (e-mail khossner@agsci.colostate.edu).

This study was designed to compare the effects of whole and size-fractionated bovine colostrum with bovine calf serum (BCS) on the growth and differentiation of 3T3-L1 fibroblasts. High (HMW) and low (LMW)-molecular-mass ultrafiltrate fractions of colostrum were prepared from defatted colostrum (COL) by diafiltration through membranes with a molecular-mass cut-off of 30 kDa. Incorporation of [³H]thymidine into the cells was used as a reflection of DNA synthesis/cell proliferation. The growth-promoting activity of LMW was 2.3- and 2.5-fold higher than COL and HMW, respectively (P < 0.05), and 185 µg/ml LMW stimulated cell proliferation equivalent to 10% BCS. Although insulin-like growth factor (IGF)-I, IGF-II and platelet-derived growth factor AB stimulated 3T3-L1 cells, antibodies to these factors did not inhibit the LMW effects. The LMW fraction was about twice as effective as COL and HMW in stimulating differentiation of the cells into adipocytes, but maximal differentiation was only 60% of that seen with 10% fetal bovine serum (FBS). Treatment with COL, HMW, IGF-I and insulin induced peroxisome-proliferator-activated receptor g RNA, but levels were about half of that with 10% FBS treatment and LMW induction was 80% of FBS. Low amounts of leptin mRNA were detected in adipocytes and abundance did not differ between treatments with BCS, hormones or COL fractions. This study showed that bovine colostral LMW stimulated the growth and differentiation of 3T3-L1 preadipocytes and may be a useful serum substitute to support the growth of these cells.

Received 18 June 2002/2 August 2002; accepted 2 September 2002

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