Hair Follicle Source Of Mesenchymal Stem Cells In Vitro Three-dimensional And Flat Culture Method Of Experimental Study

Stem cells are the body of a self-renewing, highly proliferative and multilineagedifferentiation potential, cell groups, under certain conditions, cells can differentiate into avariety of functions. Stem cell regenerative medicine primordia can be used for tissue repair,organ regeneration, gene therapy research and clinical treatment. The more in-depth researchof embryonic stem cells, bone marrow mesenchymal stem cells, umbilical cord blood stemcells, mesenchymal stem cells, hair follicles. Mesenchymal stem cells, hair follicles betweennot only self-renewal and directed differentiation of fat, bone, cartilage cells, moreimportantly, the hair follicle mesenchymal stem cells express embryo stem cell markers(SOX2, and Nanog). Rich source of hair follicles, convenient access to harvest are not subjectto age restrictions, and almost does not do any harm to the body, can also be repeated toobtain, and immunogenicity of the ground, tissue repair, organ reconstruction and genetherapy is an important source of seed cells. Requires a lot of seed cells for regenerativemedicine research and treatment, traditional flat culture it is difficult in a short period of timeto obtain a large number of seed cells, and the application of micro-carrier, three-dimensionalculture in bioreactor technology can solve this problem. Microcarrier surface area/volume (S/V), per unit volume of culture fluid cell yields; dimensional culture of part-suspension andadherent cultured advantages of simplifying the detection and control of environmentalfactors of cell growth, good reproducibility, low cost, labor intensity. Dimensional culture canbe a lot of hair follicle mesenchymal stem cells, however, its growth characteristics, biologicalcharacteristics unknown.The purpose of this experimental study is to establish vitro amplification of the human hairfollicle mesenchymal stem cells (hHF-MSCs) bioreactor technology for regenerativemedicine research and clinical treatment of undifferentiated, a large number of seed cells.Our the extractor adult hair, tissue culture, and the successful hHF-of MSCs. We will gethHF-MSCs successful vaccination to the carrier by acridine orange staining and confocalmicroscopy scanning, can be observed hHF-MSCs attached to the carrier surface and stretchfusiform. hHF-MSCs inoculation1000/cm~2or500/cm~2density to the carrier to enter thelogarithmic growth phase, proliferation to15d when cells enter the platform of the cells in theinoculation of the first3d; plane train hHF-MSCs6days into the logarithmic growing season.Mesenchymal stem cell surface markers of CD90and of CD105immunofluorescence staining,two kinds of ways to nurture hHF-MSCs were positive for expression; flow cytometryanalysis: three-dimensional to cultivate hHF-of MSCs of CD90of CD105expression rateswere80.25%,45.36%, flat culture hHF-MSCs CD90, CD105expression rates were85.65%,43.88%; these two kinds of ways to nurture hHF-MSCs after adipogenic, oil red O staining, lipid droplets red; after osteogenic induction, Alizarin red staining showed that shows thatcalcium deposition and calcium nodules were stained red.This study has successfully to obtain hHF-of MSCs, the establishment of in vitro amplifiedhHF-MSCs bioreactor technology, application of the technology culture hHF-MSCs MSCssurface markers remain, and multilineage differentiation potential, this is the post-to establishhHF-of MSCs banks, construction of tissue engineered organs, transplantation of tissues andorgans to repair lesions, as a gene therapy target cells and other experimental and clinicalapplication provides a rich source of seed cells.