Basic & Clinical Medicine ›› 2013, Vol. 33 ›› Issue (11): 1418-1421.
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Abstract: Objective To improve the methods for primary culture and purification of type 2 diabetic human keratinocytes in serum-free medium in vitro. Methods Type 2 diabetic human keratinocytes was isolated by 2 step digestion of neutral protease and trypsin, cultured in serum-free medium and purified by two-step subculture, the keratinocytes were observed using inverted phase contrast microscope, the growth curve was obtained by the WST-8 assay and phenotype and purity were analyzed by flow cytometry. Results Type 2 diabetic human keratinocytes in serum-free medium showed typical epithelial morphology, the growth curve of the keratinocyte at passage 3 is S-shaped, the average doubling time is 36.9 hours during logarithmic growth phase and the keratin-positive rate was 98.9%. Conclusion The methods for culture of high-purity type 2 diabetic human keratinocytes in serum-free media have been established.
Key words: keratinocytes, type 2 diabetes, primary culture
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URL: http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/
http://journal11.magtechjournal.com/Jwk_jcyxylc/EN/Y2013/V33/I11/1418