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  • Isolation and characterization of hair follicle stem cells from Arbas Cashmere goat.

Isolation and characterization of hair follicle stem cells from Arbas Cashmere goat.

Cytotechnology (2016-05-20)
Nimantana He, Zhenguo Dong, Li Tao, Sirguleng Zhao, Shorgan Bou, Dongjun Liu
ABSTRACT

In this study, highly purified hair follicle stem cells from Arbas Cashmere goat (gHFSCs) were isolated using enzyme digestion and adhesion to type IV collagen. The biological characteristics of the gHFSCs were identified by morphological observation, growth curve, markers assay and differentiation in vitro. The gHFSCs were in small cell size with typical cobblestone morphology, good adhesion and high refractive index. Immunocytochemistry staining showed the cells were expressing Krt15, Krt19, CD34, Itgβ1 and Krt14. Cell growth curve indicated that cultured gHFSCs had strong proliferation ability. Krt14 and CD34 were high expressed at the mRNA level, respectively, 39.68 and 24.37 times of the Cashmere goat keratinocytes, and krt15 expression was 5.62 times and itgβ1 expression was 1.81 times higher (p < 0.01). Western blot detected the expression of all the above markers. After osteogenic induction, the cells were positive for Von Kossa staining and expressed Osteocalcin. Sulfated proteoglycans in cartilaginous matrices were positively stained by Alcian blue after chondrogenic induction and COL2A1 was expressed. In myogenic induction, Hoechst 33342 staining evidenced cytoplasm fusion and positive expression of MyoG was detected by immunocytochemistry.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Insulin solution from bovine pancreas, BioReagent, 10 mg/mL insulin in 25  mM HEPES, pH 8.2, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Dispase® I, protease
Sigma-Aldrich
L-Ascorbic acid, reagent grade, crystalline
Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
Collagen from human placenta, Bornstein and Traub Type IV, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Epidermal Growth Factor from murine submaxillary gland, lyophilized powder, BioReagent, suitable for cell culture