一种有效区分冻融 3T3-L1 细胞方法的比较。
A comparison of methods for effective differentiation of the frozen-thawed 3T3-L1 cells.
机构信息
Shandong Provincial Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China.
Xiajin First Middle School of Shandong Province, Dezhou, 253200, China.
出版信息
Anal Biochem. 2019 Mar 1;568:57-64. doi: 10.1016/j.ab.2018.12.020. Epub 2018 Dec 27.
The differentiation efficiency of 3T3-L1 preadipocytes is an essential factor affecting studies on cellular mechanisms associated with obesity, diabetes, and related disorders. Differentiation of 3T3-L1 cells is commonly induced by an adipogenic cocktail containing insulin, dexamethasone (DEX), and 3-isobutyl-1-methylxanthine (IBMX). However, 3T3-L1 cells after freezing and thawing for many times always have a low differentiation efficiency. To solve this problem, we compared the differentiation efficiency of six commonly used adipogenic cocktails and protocols published in 2017. On this basis, we further compared 18 adipogenic cocktails with 2 μM rosiglitazone added and/or with a prolonged treatment with IBMX. The results revealed that the adipogenic cocktail containing 0.5 mM IBMX, 1 μM DEX, and 10 μg/mL insulin was the most effective for the frozen-thawed 3T3-L1 cells differentiation. Rosiglitazone, and IBMX under a prolonged treatment, could improve the differentiation efficiency of the frozen-thawed 3T3-L1 cells. However, the effect was closely related to concentrations of agents in the adipogenic cocktails.
3T3-L1 前脂肪细胞的分化效率是影响肥胖症、糖尿病和相关疾病相关细胞机制研究的一个重要因素。3T3-L1 细胞的分化通常通过含有胰岛素、地塞米松（DEX）和 3-异丁基-1-甲基黄嘌呤（IBMX）的脂肪生成鸡尾酒诱导。然而，经过多次冷冻和解冻的 3T3-L1 细胞的分化效率总是很低。为了解决这个问题，我们比较了 2017 年发表的六种常用的脂肪生成鸡尾酒和方案的分化效率。在此基础上，我们进一步比较了添加 2μM 罗格列酮和/或延长 IBMX 处理时间的 18 种脂肪生成鸡尾酒。结果表明，含有 0.5mM IBMX、1μM DEX 和 10μg/mL 胰岛素的脂肪生成鸡尾酒对冷冻解冻的 3T3-L1 细胞分化最有效。罗格列酮和 IBMX 的长时间处理可以提高冷冻解冻的 3T3-L1 细胞的分化效率。然而，其效果与脂肪生成鸡尾酒中试剂的浓度密切相关。
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