| Objectives:To simulate in vivo microenvironment better and to explore the regulation capacities of microencapsulated osteoblasts in ex vivo oriented differentiation of bone marrow mesenchymal stem cells (BM MSCs), BM MSCs derived from SD mouse were cocultured with osteoblasts microencapsulated in alginate-poly-lysine-alginate (APA) microcapsules in this study.Methods:Immunophenotyping of BM MSCs (P5) was analyzed by flow cytometry. And osteogenic and adipogenic induction were assessed to prove multilineage differentiation in BM MSCs used for this study. SD mouse osteoblasts (passage 3) were microencapsulated in APA microcapsules with the diameter of about 400μm and then were cocultured with BM MSCs at the cell proportion of about 10:1 (osteoblasts:MSCs) in culture plates with serum-free medium. After coculture, alkaline phosphatase (ALP) and von Kossa staining were performed to assess the osteogenic differentiation of BM MSCs.Results:BM MSCs expressed CD45- and CD29+ and had multilineage differentiation capacities. After coculture with microencapsulated osteo-blasts for several weeks, the quantitative and qualitative determination of ALP and calcium deposit indicated that BM MSCs could be induced to osteoblasts by osteoblasts encapsulated in APA microcapsules.Conclusion:Microcapsule can separate BM MSCs from osteoblasts so that the immunoreaction between these two types of cells can be avoided in this coculture system. And microencapsulated osteoblasts can effectively regulate the ex vivo oriented differentiation of BM MSCs through secreting some important soluble growth factors. This study indicated that microcapsule may be a novel coculture protocol to control ex vivo oriented differentiation of stem cells and microencapsulated coculture system will provide a novel method for MSC oriented differentiation in bone tissue engineering. |
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