I would like do some research on melanogenesis in hair follicle melanocytes. However, most of the melanoma cell line are derived from skin.
I found an article describing isolation of melanocytes from Hair follicle:
Hair follicle melanocytes (HFM) were established from normal human haired scalp tissue, obtained from 7 females (43–59 years, mean 48 years, hair color light brown to black), after elective plastic surgery with informed consent and with local ethics committee approval. All cell culture reagents were obtained from Invitrogen Ltd. (Paisley, Scotland) unless otherwise stated. HFM cultures were established using described protocols (31,32). Contaminating fibroblasts, when present in HFM cultures, were removed with 150 μg/ml geneticin sulfate (G418) in cycles of 48 h (33). Hair follicle keratinocytes (HFK) were separated from HFM cultures by differential trypsinization. The identity of isolated cells was confirmed by immunophenotyping at passage 1 with the melanocyte lineage-specific marker NKI/beteb against glycoprotein100 (gp100) (Monosan, Netherlands) (34).
Kauser, S., Slominski, A., Wei, E. T., & Tobin, D. J. (2006). Modulation of the human hair follicle pigmentary unit by corticotropin-releasing hormone and urocortin peptides. The FASEB journal, 20(7), 882-895.
My questions are
1. Is there any established hair follicle melanocytes cell lines available to be ordered?
2. Anyone have tried to use above protocols? is it easy to perform?
2. Is there any other methods to isolate hair follicle melanocytes from primary hair bulbs?
HI Alex,
Human Immortalized Melanocytes-hTERT from ABM
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Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow II medium available in ABM, Cat. No.TM002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO2), 5%.
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Real Time PCR was used to quantify hTERT gene expression in immortalized cell line.
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Best wishes,
Ilya
Ilya Tsyrlov, I have checked it. The source organ is skin. Is it from hair follicle?
It is probable that the melanocytes in hair follicles are interchangeable with those in the interfollicular skin. In people treated successfully for vitiligo, the skin can be repigmented by melanocytes that migrate out from the follicles. The follicular microenvironment may make a difference to the cell physiology, but by making a pure cell culture you lose that environment anyway. So probably there is not much point in trying to get the specific melanocytes out of hairs.
I am pretty sure that nobody has immortalized any human hairbulb melanocytes. Only a couple of groups have ever even cultured these cells.
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