Ficoll density gradient sedimentation isolation of pelage hair follicle mesenchymal stem cells from adult mouse back skin: A novel methods for hair follicle mesenchymal stem cells isolation.

Abstract:

Background: Hair follicle mesenchymal stem cells(HF-MSC) has great potential for cell therapy. Traditional method to isolate whisker HF-MSC is time consuming and few in cell numbers. How to quickly and conveniently obtain a large number of HF-MSC for experimental research is a problem worth exploring.

Methods: Two steps Ficoll Density Gradient Sedimentation(FDGS) were performed to isolated pelage HF-MSC from adult mice. The characteristic of the isolated cells were identified and compared with whisker HF-MSC by immunofluorescence staining, flow cytometry, three-lineage differentiation and hair follicle reconstruction. Pelage HF-MSC were injected into the dorsal skin of mice to explore its role in promoting hair growth. The cells and exosomes distribution were located by immunofluorescence staining.

Results: Isolated pelage HF-MSC expressed similar markers (ALP, Versican, NCAM, Nestin), showed similar growth pattern, possessed similar mesenchymal stem cells function and hair follicle induction ability as whisker HF-MSC. A large number of cells can be obtained with fewer mice compared to tranditional method. Pelage HF-MSC injection can promote hair growth. Pelage HF-MSC were distributed surround the host hair follicle, secreting exosomes that integrated into host hair follicle and located in dermal papillae and hair matrix region.

Conclusion: A large number of Pelage HF-MSC can be isolated by FDGS, which can promote hair growth by secreting exosomes which targeted the dermal papilla and hair matrix region of host hair follicle.