Merck
  • Home
  • Search Results
  • 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
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MCDB 201 Medium, With trace elements, L-glutamine and 30 mM HEPES, powder, suitable for cell culture
Sigma-Aldrich
ITS Liquid Media Supplement (100×), liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Linoleic Acid-Albumin from bovine serum albumin, liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Cholera Toxin from Vibrio cholerae, ≥90% (SDS-PAGE), lyophilized powder
Sigma-Aldrich
Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Phorbol 12-myristate 13-acetate, ≥99% (TLC), film or powder
Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Sigma-Aldrich
Monoclonal Anti-Microphthalmia antibody produced in mouse, clone C5, purified immunoglobulin
Sigma-Aldrich
Gelatin from bovine skin, Type B, powder, BioReagent, suitable for cell culture