Preliminary study on human bone marrow mesenchymal stem cells differentiation into epithelial-like cells
HOU Guang-hui, YE Nan, LIU Xiao-hao, ZHANG Ze-hua
Published 2010-08-11
Cite as Chin J Ophthalmol, 2010,46(08): 719-724. DOI: 10.3760/cma.j.issn.0412-4081.2010.08.010
Abstract
Objective To investigate the differentiation potential of human bone marrow mesenchymal stem cells (hMSC) inducing into epithelial-like cells, even comeal epithelial-like cells, and to discuss the plasticity that make hMSC the seed cells used in comeal tissue engineering. Methods hMSC were isolated and purified by density gradient centrifugation combined with an attachment culture method and passaged in vitro. hMSC were identified by flow cytometry. The passaged hMSC were planted on fresh pig comeal Bowman's membrane. The expression of CK12, ABCG2 and CK19 in hMSC was identified by immunofluorescence staining. We used in vitro method to obtain a multilayer culture of hMSC. When hMSC formed a monolayer, the cells were inserted to Millicell culture and grew into multilauers by using the airlifting cultivation methodology. Four weeks later, after fixed and dehydrated, the hMSC were observed under the light microscope after hemotoxylin and eosin (HE) and immunohistochemistry staining. Results hMSC could be cultured, expanded in vitro, and showed great potential of proliferation. The result of flow cytometry showed that the positive staining percentage was 0.06% for CD45, 0.41% for CD34, 86.43% for CD44, 85.72% for CD29 and 90.72% for CD105. This indicated that hMSC expressed CD44, CD29, CD105 but not CD45 and CD34. After four weeks induction, part of hMSC expressed CK12 and CK19 but not ABCC2. In the in vitro stratification, HE and immunohistochemical staining showed that there were one or two layers epithelial-like cells, even corneal epithelial-like cells after using the air-lifting cultivation. Conclusions This study suggests that hMSC have the potential to differentiate into epithelial cells, even corneal epithelial cells. hMSC could be the option of cells used to reconstruct the comeal epithelium by tissue engineering technology.
Key words:
Mesenchymal stem cells; Cell differentiation; Comeal; Epithelioid cells; Tissue engineering
Contributor Information
HOU Guang-hui
Department of Ophthalmology, the People's Hospital of Zhuhai city, Zhuhai 519000,Guangdong 519000, China
YE Nan
Department of Ophthalmology, the People's Hospital of Zhuhai city, Zhuhai 519000,Guangdong 519000, China
LIU Xiao-hao
Department of Ophthalmology, the People's Hospital of Zhuhai city, Zhuhai 519000,Guangdong 519000, China
ZHANG Ze-hua
Department of Ophthalmology, the People's Hospital of Zhuhai city, Zhuhai 519000,Guangdong 519000, China