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Induction of Human Umbilical Cord Mesenchymal Stem Cells into Type II Alveolar Epithelial Cells In Vitro
Liu Jiang1, Peng Danyi1, Gou Hao1, Wu Xian1, Si Daozhu1, Niu Chao2, Tian Daiyin2, Liu Enmei2, Zou Lin3, Fu Zhou1,2*
1Department of Pediatric Research Institute, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Engineering Research Center of Stem Cell Therapy, Chongqing 400014, China; 2Respiratory Center of Children’s Hospital of Chongqing Medical University, Chongqing 400014, China; 3Center for Clinical Molecular Medicine, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China
Abstract: This study aimed at the directional differentiation of human umbilical cord mesenchymal stem cells (hUC-MSC) into type II alveolar epithelial cells (AEC2s) induced by a conditional medium with growth factors， so as to provide the foundation for further study on the therapeutic effects of hUC-MSCs-derived AEC2s in pulmonary diseases. hUC-MSCs cultured in vitro were divided into test group， in which cells were cultured in small air way epithelial cells growth basal medium for 2 days and then growth factors were added for the induction culture; and control group， in which cells were cultured in DMEM/F12 culture medium containing 20% fetal bovine serum. 14 days later， the cellular morphology was observed and recorded with an inverted microscope. The protein level of pro-surfactant protein C (proSP-C) was detected by Western blot， immumofluorescence method and flow cytometry， respectively. The content of surfactant protein C (SP-C) secreted by the cells in medium supernatant was detected by ELISA. The formation of lamellar body in cells was observed by a transmission electron microscope (TEM). The results indicated that test group cells’ shape transformed from long fusiform to polygon after the induction， while control group cells’ shape changed little. The protein level of proSP-C was detectable in test group cells. Secreted SP-C was detectable in medium supernatant. Lamellar body formation was observed under TEM. The proSP-C， SP-C secretion or lamellar body was not detectable in control group cells. It concluded that in vitro directional induction culture promoted the differentiation of hUC-MSCs into AEC2s. Hopefully， a large scale of hUC-MSCs-derived AEC2s could be obtained through this method.
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