Keratinocyte serum-free medium promotes the growth of rat hair follicle stem cells

doi:10.3969/j.issn.2095-4344.2013.45.015

Abstract

Abstract:

BACKGROUND: The hair follicle stem cells are usual cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10% fetal bovine serum. Recently, keratinocyte serum-free medium has been used to culture hair follicle stem cells.
OBJECTIVE: To observe the effects of three different culture media on the proliferation and purity of hair follicle stem cells.
METHODS: After sterling, the skin of the rat’s beard was cut off, and the vibrissa follicles were dissected under a stereocroscope. The follicles were digested by Dispase Ⅱ firstly and by the mixture of trypsin and EDTA secondly. Cell suspension was divided into three pieces based on the average number of cells and respectively cultured by keratinocyte serum-free medium, Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10% fetal bovine serum and keratinocyte serum-free medium containing 10% fetal bovine serum. Hair follicle stem cells were selected by rapid adherence on collagen type Ⅳ, and passaged.
RESULTS AND CONCLUSION: Cultured hair follicle stem cells spread to the third generation, and trypan blue assay showed there was no difference in cell survival rate among the three groups (P > 0.05). Cell Counting Kit 8 assay showed that the cells grew slowly in the three groups in the first 2 days. At day 4, the hair follicle stem cells grew into the growth logarithmic phase, and the proliferative activity was ranked as follows: keratinocyte serum-free medium+10% fetal bovine serum > Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum > keratinocyte serum-free medium with a significant difference between the three groups (P < 0.05). Flow cytometric examination showed the expression of CD34 in the keratinocyte serum-free medium+10% fetal bovine serum group was lower than that in the keratinocyte serum free medium group (P < 0.05). The expressions of CD34, CK15 and β1-integrin (CD29) in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum were lower than those in the other two groups (P < 0.05). These findings indicate that we can obtain higher purity hair follicle stem cells in the keratinocyte serum-free medium+10% fetal bovine serum than in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum.

Key words: stem cells, vibrissae, hair follicle, culture media, serum-free, proliferation(2012ZZGC05)*

CLC Number:

R394.2

Zhang Zhi-qiang, Wang Yu-jie, Muratrixat, Li Jia. Keratinocyte serum-free medium promotes the growth of rat hair follicle stem cells[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2013.45.015.

Figures/Tables 3

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