Lee Seong-Ho, Hossner Kim L
Department of Animal Sciences, Colorado State University, Fort Collins, CO 80523-1171, USA.
Biotechnol Appl Biochem. 2002 Dec;36(3):205-12. doi: 10.1042/ba20020060.
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 [(3)H]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 microg/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 gamma 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.
本研究旨在比较全脂及不同分子量分级的牛初乳与牛犊血清(BCS)对3T3-L1成纤维细胞生长和分化的影响。通过使用截留分子量为30 kDa的膜进行渗滤,从脱脂初乳(COL)中制备了高(HMW)和低(LMW)分子量的初乳超滤级分。将[³H]胸腺嘧啶掺入细胞中作为DNA合成/细胞增殖的反映。LMW的促生长活性分别比COL和HMW高2.3倍和2.5倍(P<0.05),185 μg/ml的LMW刺激细胞增殖的效果与10% BCS相当。尽管胰岛素样生长因子(IGF)-I、IGF-II和血小板衍生生长因子AB可刺激3T3-L1细胞,但针对这些因子的抗体并不抑制LMW的作用。LMW级分在刺激细胞分化为脂肪细胞方面的效果约为COL和HMW的两倍,但最大分化程度仅为10%胎牛血清(FBS)的60%。用COL、HMW、IGF-I和胰岛素处理可诱导过氧化物酶体增殖物激活受体γ RNA,但水平约为10% FBS处理的一半,LMW诱导水平为FBS的80%。在脂肪细胞中检测到少量瘦素mRNA,且BCS、激素或COL级分处理之间的丰度没有差异。本研究表明,牛初乳LMW可刺激3T3-L1前脂肪细胞的生长和分化,可能是支持这些细胞生长的有用血清替代品。