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Hyaluronic Acid Gel Induction MDSC Differentiation for Chondroid Cells

Author: ShenZhaoLiang
Tutor: MeiXiFan
School: Liaoning Medical
Course: Surgery
Keywords: muscle-derived stem cells cartilage-like cells cell differentiation culture in vitro cartilage cell culture medium
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


ObjectiveTo determine the concentration of hyaluronic acid gel fits to the growth of rabbit muscle-derived stem cells. The establishment of rabbit articular chondrocytes isolated and cultured by new methods. The cartilage cells as the induction factors, induced muscle-derived stem cells differentiate into cartilage-like cells in the hyaluronic acid and the induced cells were identified. To discussion the feasibility of mixed culture muscle-derived stem cells and chondrocytes to chondrogenesis in vitro.MethodsIsolated culture and amplification of passage of MDSCs and articular chondrocytes from rabbit. They mixed by a certain percentage. Then they inoculated in 5% hyaluronic acid gel with the density of 6×1010/L as co-culture group. The same density of MDSCs as negative control group, and the same density of articular chondrocytes as positive control group. Observed with inverted microscope. They detected by toluidine blue staining andⅡcollagen immunocytochemistry after 10 days. RT-PCR detected the mRNA expression ofⅡcollagen and aggrecan.ResultsThe well growth state of MDSCs is cultured in different concentrations of hyaluronic acid gel. The method of digestion with mix enzyme and cultured in vitro can acquire the large quantity of pure isolated articular cartilage cells. The chondrocyte were verified by the positive results of toluidine blue staining and collagenⅡthrough immunocytochemistry. the muscle-derived stem cells were inducted in the hyaluronic acid gel by cartilage cells, In co-culture group, the form of MDSCs changed from fusiform to multilateral polygon after 5 days. After 10 days, toluidine blue staining andⅡcollagen immunocytochemistry of cells were the same to positive control group, RT-PCR detected the mRNA expression ofⅡcollagen and aggrecan were positive. They were not differentiated to chondrocytes in negative control group.ConclusionsCartilage microenvironment plays an important role in MDSCs chondrocyte differentiation. Chondrocytes can effective induce MDSCs to cartilage-like cells. Hyaluronic acid gel may be a good vector in cartilage tissue engineering.

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