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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
ZUSAMMENFASSUNG

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).

MATERIALIEN
Produktnummer
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Produktbeschreibung

Sigma-Aldrich
Dulbecco Modifiziertes Eagle-Medium – hoher Glucosegehalt, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
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Dexamethason, powder, BioReagent, suitable for cell culture, ≥97%
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Phorbol-12-myristat-13-acetat, ≥99% (TLC), film or powder
Sigma-Aldrich
Choleratoxin aus Vibrio cholerae, ≥90% (SDS-PAGE), lyophilized powder
Sigma-Aldrich
ITS Flüssigmedien-Zusatz (100×), liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
L-Ascorbinsäure, powder, suitable for cell culture, γ-irradiated
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Gelatine aus Rinderhaut, Type B, powder, BioReagent, suitable for cell culture
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Linoleic Acid-Albumin from bovine serum albumin, liquid, sterile-filtered, BioReagent, suitable for cell culture
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Medium MCDB 201, With trace elements, L-glutamine and 30 mM HEPES, powder, suitable for cell culture
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Anti-TRP1/TYRP1-Antikörper, Klon TA99, azidfrei, clone TA99, 1 mg/mL, from mouse
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Monoclonal Anti-Microphthalmia antibody produced in mouse, clone C5, purified immunoglobulin