2. Academic Validation
  3. Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues

Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues

  • J Vis Exp. 2020 Jul 8;(161). doi: 10.3791/61226.
Chang Liu 1,  Shuangshuang Wang 1,  Man Liu 2,  Fuxiang Bai 1,  Zhihong Chen 3,  Ping Wang 4,  Jun Mi 5,  Xunwei Wu 6
Affiliations

Affiliations

  • 1 Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration.
  • 2 Shijiazhuang Shimen Experimental School.
  • 3 Qilu Children's Hospital of Shandong University.
  • 4 Qilu Hospital of Shandong University.
  • 5 Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration; Shenzhen Research Institute of Shandong University; mijun1984@126.com.
  • 6 Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration; xunwei_2006@hotmail.com.
PMID: 32716369 DOI: 10.3791/61226
Abstract

The isolation and culture of primary melanocytes from skin tissues is very important for biological research and has been widely used for clinical applications. Isolating primary melanocytes from skin tissues by the conventional method usually takes about 3 to 4 weeks to passage sufficiently. More importantly, the tissues used are usually newborn foreskins and it is still a challenge to efficiently isolate primary melanocytes from adult tissues. We recently developed a new isolation method for melanocytes that adds Y-27632, a Rho kinase inhibitor, to the initial culture medium for 48 h. Compared with the conventional protocol, this new method dramatically increases the yield of melanocytes and shortens the time required to isolate melanocytes from foreskin tissues. We now describe this new method in more detail using adult epidermis to efficiently culture primary melanocytes. Importantly, we show that melanocytes obtained from adult tissues prepared by this new method can function normally. This new protocol will significantly benefit studies of pigmentation defects and melanomas using primary melanocytes prepared from easily accessed adult skin tissues.

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