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一、四川農(nóng)業(yè)大學農(nóng)場動物遺傳資源探索與創(chuàng)新重點實驗室，使用美國zeta life胎牛血清成功培養(yǎng)兔前脂肪細胞(preadipocytes)發(fā)表文章已見刊；

二、培養(yǎng)條件：

培養(yǎng)細胞名稱：兔前脂肪細胞(preadipocytes)

培養(yǎng)體系：6孔板

培養(yǎng)細胞數(shù)量：6 × 105

三、用zeta life胎牛血清培養(yǎng)前脂肪細胞(preadipocytes)發(fā)表文章部分內容：

Isolation and induction of rabbit preadipocytes Visceral preadipocytes were isolated from perirenal adipose tissue of newborn New Zealand rabbits under sterile conditions. Briefly, adipose tissues were digested with 0.01% collagenase I (Gibco, Carlsbad, CA, USA). Then, preadipocytes were seeded into 6-well plates at a density of 6 × 105 cells per plate in complete medium (DME/F12, supplemented with 10% fetal bovine serum [FBS]) (DME/F12 was obtained Gibco, Carlsbad, CA, USA; fetal bovine serumwas from Zeta life, Menlo Park, CA, USA), and preadipocyteswere cultured in a humidified incubator at 37 °C and 5% CO2. Upon reaching confluency (set to day 0), cells were induced by the addition of differentiation medium I (DME/F12 with 1 μM dexamethasone, 500 μM 1-methy1-3-iosbutylxanthine, 1.7 μM insulin, 10% FBS) (dexamethasone, 1-methy1-3- iosbutylxanthine, and insulin were from Solarbio, Beijing, China) for 72 h (day 3). Next, cells were cultured with differentiation medium II (DME/F12, 1.7 μM insulin, 10% FBS) for an additional 72 h and further cultured in complete medium until day 9 (day 9). To identify lipid accumulation in cells and obtain mature adipocytes, the accumulation of lipid droplets was measured by Oil Red O staining. Based on three different phenotypes of lipid accumulation (almost no lipid droplets, a small amount of ring-shaped lipid droplets, and a cluster of bigger lipid droplets), a total of nine cell samples were collected, and each interval time point contained three biological replicates. RT-qPCR was performed to determine the mRNA expression levels of adipogenic marker genes, including PPARG, CEBPA, and FABP4 (primer sequences are shown in Table S1), and the 2-ΔΔCt method was used to analyze the relative expression. The housekeeping gene GAPDH served as control.

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