zeta life胎牛血清培養(yǎng)前脂肪細(xì)胞發(fā)表文章已見刊

一、四川農(nóng)業(yè)大學(xué)農(nóng)場(chǎng)動(dòng)物遺傳資源探索與創(chuàng)新重點(diǎn)實(shí)驗(yàn)室，使用美國(guó)zeta life胎牛血清成功培養(yǎng)兔前脂肪細(xì)胞(preadipocytes)發(fā)表文章已見刊；

二、培養(yǎng)條件：

培養(yǎng)細(xì)胞名稱：兔前脂肪細(xì)胞(preadipocytes)

培養(yǎng)體系：6孔板

培養(yǎng)細(xì)胞數(shù)量：6 × 105

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

Isolation and induction of rabbit preadipocytes Visceral preadipocytes were isolated from perirenal adipose tissue of newborn New Zealand rabbits under sterile condi[1]tions. 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, sup[1]plemented 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 differ[1]entiation medium II (DME/F12, 1.7 μM insulin, 10% FBS) for an additional 72 h and further cultured in complete medi[1]um until day 9 (day 9). To identify lipid accumulation in cells and obtain mature adipocytes, the accumulation of lipid drop[1]lets 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, in[1]cluding PPARG, CEBPA, and FABP4 (primer sequences are shown in Table S1), and the 2-ΔΔCt method was used to ana[1]lyze the relative expression. The housekeeping gene GAPDH served as control.

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