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上海創(chuàng)凌生物科技有限公司>技術(shù)文章>美國zeta life 胎牛血清 成功培養(yǎng)前脂肪細(xì)胞發(fā)表文章已見刊

技術(shù)文章

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

閱讀:1404          發(fā)布時間:2020-6-15

一、四川農(nóng)業(yè)大學(xué)農(nóng)場動物遺傳資源探索與創(chuàng)新重點(diǎn)實(shí)驗(yàn)室,使用美國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 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.

zeta life胎牛血清信息如下

產(chǎn)品名稱

貨號

規(guī)格

數(shù)量

Fetal Bovine Serum France Origin

Z7186FBS-500

500ml

1

Fetal Bovine Serum France Origin 

Z7186FBS-100

100ml

1

Fetal Bovine Serum ,South America Origin

Z7181FBS-500

500ml

1

Fetal Bovine Serum ,South America Origin 

Z7181FBS-100

100ml

1

Fetal Bovine Serum (South America Origin) Ultra-low Endotoxin

Z7180FBS-500

500ml

1

Fetal Bovine Serum (South America Origin) Ultra-low Endotoxin

Z7180FBS-100

100ml

1

Fetal Bovine Serum (Australia Origin)

Z7010FBS-500

500ml

1

Fetal Bovine Serum (Australia Origin)

Z7010FBS-100

100ml

1

Fetal Bovine Serum ,South America Origin

Z7185FBS-500

500ml

1

Fetal Bovine Serum ,South America Origin

Z7185FBS-100

100ml

1

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