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| Optimization of Dermal Papilla Cell Cultivation Condition In vitro in Cashmere Goat (Capra hircus) |
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Abstract Dermal papilla cells (DPCs) are specialized mesenchymal-originated fibroblasts with prominent ability to induce hair follicle morphogenesis and development in mammals. They are situated at the bottom of hair follicle and direct hair follicle remodeling by sending various growth factors to follicular keratinocytes. Isolation and culture of DPCs from hair follicle is laborious and time-consuming job, thus present study was designed to optimize the culture condition of goat DPCs in order to acquire enough DPCs for other studies. In this study, DP was separated and cultured from goat hair follicle under stereoscope by microdissection, and confirmed the identity of cultured cell by using antibodies specific to their marker genes α-smooth muscle actin and CD133/prominin-1, then cell counting, (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay, alkaline phosphatas analysis and other methods were used to compare the combination of serum concentration and medium types. The results indicated that (1) goat DPCs shared the expression of specific markers as previously reported, and they were successfully cultured in four tested culture medium supplemented with different proportions of serum respectively; (2) Growth of goat DPCs depends on the nutritional supplement: the higher proportion of serum added in culture medium resulted faster proliferation rate and stronger ability to migrate and induce hair growth; (3) Advanced medium does contain effective factors to maintain or strengthen goat DPCs proliferation and inducing ability, thus it is more suitable for propagating goat DPCs than conventional medium, otherwise when the serum addition proportion is 10%, the growth and proliferation of goat DPCs cultured advanced medium are significantly higher than that in conventional media (P＜0.05); (4) the effect of Advanced media added with 4%~6% serum for gaot DPCs propagation is as good as the normal medium added with 10% serum (P＞0.05). In conclusion, we successfully cultured goat DPCs, identified them and proved that Advanced medium are more suitable for propagation of them in vitro than conventional medium.
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Received: 14 June 2017
Published: 04 February 2018
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| Fund:;Genetically modified organisms breeding major projects |
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