Human umbilical cord blood stem cells differentiate into nasal ciliated epithelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Damage to nasal ciliated epithelial cels can lead to a severe injury in nasal biological function. Compared with other adult stem cels, human umbilical cord blood stem cels have better differentiation potential. OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cels differentiating into nasal ciliated epithelial cels through in vitro culture and induction techniques. METHODS:Normal and healthy umbilical cord blood samples were colected to isolate human umbilical cord blood stem cels, folowed by identification and subculturein vitro. Umbilical cord blood stem cels at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cels at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks. RESULTS AND CONCLUSION:After subculture, passage 3 umbilical cord blood stem cels that could express stem cel surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cels were visible, and the cel positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cels, but expressed strongly in the nasal ciliated epithelial cels, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionaly, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cels could differentiate into nasal epithelial cels under suitable conditions.