Fig. 2. Purification of primary melanocytes by FACS sorting from neonatal dermal suspensions of C57BL/6 mice. Primary cells were cultured from dermal suspensions of C57BL/6 neonates for a period of 10 days with TPA, bFGF and dbcAMP as described. Primary cells lacking additives for 72 hours were double labeled with PE-CD117 (anti-Kit) and APC-CD45 and separated by FACS. The CD117+CD45-population was plated and analyzed for melanocyte content using MEL-5 (anti-Tyrp1) immunofluorescence. (A) Anti-CD117 (against Kit) and anti-CD45 flow cytogram of mixed primary cultures from murine neonatal dermal suspensions of wild-type cells. The CD117⁺CD45^– population was generally 1% of the total population of cells. (B) Bright-field photomicrograph of CD117⁺CD45^– population showing prominent pigmentation and dendrites characteristic of melanocytes. (C) MEL-5 (anti-Tyrp1) immunofluorescence of sorted CD117⁺CD45^– cells. (D) DAPI staining of cells shown in C.