Study on the osteogenic effects of type II arabinogalactan on the human mesenchymal stem cells

• Main Authors: Bo-Zih Shu, 許博諮
• Other Authors: Huey-Chun Huang
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/v88u4n

碩士 === 中國醫藥大學 === 醫學檢驗生物技術學系碩士班 === 102 === Human mesenchymal stem cells (hMSC) are self-renewing precursor cells that can be expanded in vitro and differentiated towards osteogenic, chondrogenic or adipogenic lineages. However, the limit number and inefficient hMSCs differentiation are the major barriers in hMSC-based therapy. It would be extremely beneficial if a naturally occurring agent could be identified that could induce hMSCs to undergo specific lineage differentiation. In the present study, we investigated the potential ability of type II arabinogalactan to enhance osteogenesis in hMSC cells and to determine the mechanism of action. By using flow cytometry, hMSCs were characterized of the specific markers CD44, CD90. hMSCs were successfully differentiated into adipocytes and osteocytes after cultivation in induction media. In addition, the expression of p53 were decreased after hMSCs differention. hMSCs differentiated into adipocytes were analyzed by Oil-Red O staining. Osteogenic differentiation was identified by the mineralization of calcium deposits using Alizarin S staining. The ERK, AKT ,PKC protein expression were detected using Western blot .The mRNA and protein expression of osteogenic marker genes (Runx2, type I collagen, osteopontin /OP, osteocalcin/OC, alkaline phosphatase) were detected using Q-PCR and Immunofluorescence staining. Our results showed that type II arabino- galactan could enhance the gene expression of osteogenic markers and mineralization of hMSC ,and the ERK, AKT ,PKC protein expression were increase in osteogenic of hMSC ,and the type II arabinogalactan enhances osteogenic differentation of hMSC via decrease of p53 expression.