Differentiation of canine bone marrow mesenchymal stem cells into epithelium-like cells: An in vitro study
- Author:
Gui-Lin HAN
1
Author Information
1. Department of Cardiothoracic Surgery
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(8):858-862
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the feasibility of inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into epithelium-like cells in vitro and the basic inducing conditions. Methods: BMSCs were obtained from healthy adult canine bone marrow aspirates and were purified by density gradient centrifugation and anchoring culture. BMSCs of second passage were cultured with 4 different media: K-SFM+EGF+BPE, DMEM+EGF, K-SFM+BPE, and DMEM+FBS. The changes in cell morphology were observed daily under phase-contrast microscopy and cell growth curves of every group were drawn by the continual cell counting. The cells were identified by immunocytochemistry staining and flow cytometer through detecting cytokeratin (CK) and epithelial membrane antigen (EMA) on the 7th and 14th day of induction. Results: A few BMSCs in K-SFM+EGF+BPE group became round or oval with processes on the 7th day of induction; more BMSCs became round or oval and their phenotypic markers were CK+ and EMA+ on the 14th day. FACS analysis showed that CK or EMA positive cells were rare in K-SFM+EGF+BPE group on the 7th day after induction (<1%), and the number of positive cells were increased (> 7%) on the 14th day. Cell morphology had no obvious change in DMEM+EGF, K-SFM+BPE, and DMEM+FBS groups during the whole culture process. Moreover, cells in DMEM+EGF, K-SFM+BPE, and DMEM+FBS group were negative for CK and EMA. Conclusion: Our experiment indicates that the combination of K-SFM, EGF and BPE can induce canine BMSCs to differentiate into epithelium-like cells in vitro.
