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  • Generation of Human iMelanocytes from Induced Pluripotent Stem Cells through a Suspension Culture System.

Generation of Human iMelanocytes from Induced Pluripotent Stem Cells through a Suspension Culture System.

STAR protocols (2020-10-29)
Li-Ping Liu, Ning-Ning Guo, Yu-Mei Li, Yun-Wen Zheng
摘要

Melanocytes, derived from neural crest cells, are involved in melanin production. This protocol describes a method to generate induced melanocytes (iMelanocytes) from human induced pluripotent stem cells (iPSCs) using a suspension culture system, which considerably improves the differentiation efficiency. The most critical parts of this protocol are the selection of a reliable iPSC line with strong potential to differentiate into melanocytes and their stemness maintenance. For complete information on the use and generation of this protocol, please refer to our Cell Reports article, Liu el al. (2019).

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杜氏改良 Eagle 培养基 - 高葡萄糖, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
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地塞米松, powder, BioReagent, suitable for cell culture, ≥97%
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佛波醇12-十四酸酯13-乙酸酯, ≥99% (TLC), film or powder
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ITS液体培养基补充剂(100x), liquid, sterile-filtered, BioReagent, suitable for cell culture
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L -抗坏血酸, powder, suitable for cell culture, γ-irradiated
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明胶 来源于牛皮, Type B, powder, BioReagent, suitable for cell culture
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亚油酸蛋白 来源于牛血清白蛋白, liquid, sterile-filtered, BioReagent, suitable for cell culture
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MCDB 201 培养基, With trace elements, L-glutamine and 30 mM HEPES, powder, suitable for cell culture
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抗-TRP1/TYRP1抗体(克隆TA99,不含叠氮化物), clone TA99, 1 mg/mL, from mouse