Osteogenic differentiation potential of microencapsulated transgenic bone marrow mesenchymal stem cells cocultured with osteoblasts

doi:10.12307/2022.565

Abstract

Abstract: BACKGROUND: With the rapid development of transgenic technology, gene therapy and microencapsulation technology are combined to form microencapsulated gene delivery technology. In addition to providing a good three-dimensional microenvironment for the growth of stem cells and ensuring the large-scale in vitro culture of stem cells, microencapsulation can also use selective permeability membranes to isolate the graft from the host immune system, and effectively avoid immune rejection during allogeneic transplantation.
OBJECTIVE: To observe proliferation activity and osteogenetic differentiation potential after coculture of microcapsuled genetically modified bone marrow mesenchymal stem cells (BMSCs) and osteoblasts.
METHODS: APA-Foxc2-BMSCs microcapsule complexes were prepared using pulsed high voltage electrostatic microcapsule preparation instrument. Acridine orange/ethidium bromide staining was conducted. Microencapsulated transgenic BMSCs and osteoblasts were cocultured. At 3 weeks after coculture, MTT assay was performed to detect cell proliferation. At 1 week, quantitative detection of alkaline phosphatase activity was performed. At 1 and 2 weeks, qualitative detection of alkaline phosphatase and von Kossa staining were conducted. At 2 weeks, western blot assay and real time PCR were performed to measure osteogenic factor expression.
RESULTS AND CONCLUSION: (1) Acridine orange/ethidium bromide staining demonstrated that 70%-80% of the cells in the microcapsule dyed green fluorescence. Under the microscope, cells did not escape from the microcapsule or stain outside the capsule. After breaking capsule, cultured BMSCs grew well again. (2) In the microencapsulated coculture group, alkaline phosphatase staining showed many tan particles in the cytoplasm. Alkaline phosphatase activity was significantly increased, and calcium matrix deposition appeared. (3) In the microencapsulated coculture group, expression of type I collagen, platelet-derived growth factor protein and mRNA significantly increased. (4) The results suggest that microcapsuled transgenic BMSCs cocultured with osteoblasts can strengthen BMSCs proliferation activity and promote BMSCs osteogenic differentiation.

Key words: bone marrow mesenchymal stem cells, microencapsulated, osteoblasts, coculture, proliferation, osteogenic differentiation

CLC Number:

You Wulin, Huang Guicheng, Wang Jianwei. Osteogenic differentiation potential of microencapsulated transgenic bone marrow mesenchymal stem cells cocultured with osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(24): 3852-3857.

Figures/Tables 11

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