Cultivation of Hair Matrix Cells from Cashmere Goat Skins and Exemplified Applications

SIMPLE SUMMARY: A large scale of sequencing data pertaining to cashmere growth on cashmere goats have not been cost-effectively used due to the lack of in vitro cellular models, especially for hair matrix cells (HMCs)—the precursors of hair-forming keratinocytes, causing an enormous waste of data resources. Herein, we successfully isolated and cultivated previously unreported HMCs from cashmere goat skins and identified them morphologically and molecularly via their distinct appearance and signature genes’ expression from spatially adjacent dermal papilla cells. Through monitoring the effects of calcium and all-trans retinoic acid on HMCs using various biological techniques, we displayed that the cells are useful models to explore unsolved issues in hair fiber growth on goats. Therefore, our present success paves the road for further utilizing currently deposited data to unveil the secrets of cashmere growth and, ultimately, improve the quantity and quality of animal fibers. ABSTRACT: A functional interpretation of filtered candidates and predicted regulatory pathways related to cashmere growth from sequencing trials needs available cell models, especially for hair matrix cells (HMCs), whose continual proliferation and differentiation result in rapid hair growth. To fulfill such goals, we herein obtained primary goat HMCs via a microdissection-based method; optimized the selection of the culture medium and coating substances for better cell maintenance; and exemplified their usefulness through examining the effects of calcium and all-trans retinoic acid (ATRA) on cells using immunoblotting, flow cytometry, and other techniques. As a result, we successfully acquired primary and passaged goat HMCs with typical keratinocyte morphology. Calcium-free RPMI (Roswell Park Memorial Institute) 1640 and MEM (minimum Eagle’s medium) outperformed normal DMEM/F12 (Dulbecco’s modified Eagle’s medium/Nutrient Mixture F-12) on long-term cell maintenance, whereas serum-free media K-SFM and EpiLife failed to support cell growth. HMCs differed molecularly and morphologically from their neighbor dermal papilla cells on expressions of feature genes, such as HOXC13, and on characteristic keratinocyte-like appearances versus fibroblast shapes, respectively. Higher calcium concentrations significantly stimulated the expression of the genes (e.g., KRT1 and IVL) involved in keratinocyte differentiation and, promoted cell proliferation. Moreover, 10(−5) M ATRA obviously boosted goat HMC expansions and changed their cell cycle distributions compared to the controls. Our study shines a light on researches exploring the mechanisms underlying the growth of cashmere.