Adipose-derived stem cells differentiate to keratocytes in vitro

[摘要] Purpose: Adipose-derived stem cells(ADSC) are an abundant population of adult stem cells with thepotential to differentiate into several specialized tissue types,including neural and neural crest-derived cells. This study sought todetermine if ADSC express keratocyte-specific phenotypic markers whencultured under conditions inducing differentiation of corneal stromalstem cells to keratocytes. Methods: Human subcutaneous adiposetissue was obtained by lipoaspiration. ADSC were isolated bycollagenase digestion and differential centrifugation. Side populationcells in ADSC were demonstrated using fluorescence-activated cellsorting after staining with Hoechst 33342. Differentiation tokeratocyte phenotype was induced in fibrin gels or as pellet cultureswith serum-free or reduced-serum media containing ascorbate.Keratocyte-specific gene expression was characterized using westernblotting, quantitative RT–PCR, and immunostaining. Results: ADSC contained a sidepopulation and exhibited differentiation to adipocytes and chondrocytesindicating adult stem-cell potential. Culture of ADSC in fibrin gels oras pellets in reduced-serum medium with ascorbate and insulin inducedexpression of keratocan, keratan sulfate, and aldehyde dehydrogenase 3family, member A1 (ALDH3A1), products highly expressed bydifferentiated keratocytes. Expression of differentiation markers wasquantitatively similar to corneal stromal stem cells and occurred inboth serum-free and serum containing media. Conclusions: ADSC cultured underkeratocyte-differentiation conditions express corneal-specific matrixcomponents. Expression of these unique keratocyte products suggeststhat ADSC can adopt a keratocyte phenotype and therefore have potentialfor use in corneal cell therapy and tissue engineering.

[学科分类] 生物化学/生物物理