Chondrgenic differentiation of human bone marrow-derived mesenchymal stem cells by novel small-molecule inducers

small molecule;TGF-β3;human bone marrow;mesenchymal stem cell;chondrogenic differentiation;중배엽 줄기세포;저분자 화합물;TGF-β3;연골분화;골수줄기세포;617

치과대학 치의과학과 ;

University:서울대학교 대학원

关键词: small molecule;    TGF-β3;    human bone marrow;    mesenchymal stem cell;    chondrogenic differentiation;    중배엽 줄기세포;    저분자 화합물;    TGF-β3;    연골분화;    골수줄기세포;    617;   

Others  :  http://s-space.snu.ac.kr/bitstream/10371/131141/1/000000021161.pdf

美国|英语

来源: Seoul National University Open Repository

【 摘 要 】

To treat cartilage defection, mesenchymal stem cells(MSCs) are considered as promising alternative cell source. Especially bone marrow derived MSCs have best characterized adult stem cells and have been used in various chondrogenic differentiation researches. Inducing chondrogenesis from human bone marrow derived MSCs is formed in combination of defined media and TGF-β3. We also found small molecules 20B09, 20F08 which have the effect to cells by high throughput screening (HTS) method. The purpose of the experiment is to confirm whether these small molecules are able to replace TGF-β3. We measured the chondrogenic differentiation ability of 20B09, 20F08 after physical triggering by centrifugation. After 7 and 14 days, three dimensional chondrogenic clusters were measured by GAG assay, RT-PCR, specical staining such as Safranin-O and Von Kossa. Furthermore, we measured collagenⅠ, Ⅱ by immunohistochemistry. Macroscopic results for 14 days were not different from TGF-β3 groups and small molecule groups. GAG assay showed that 20B09 groups were the faster enrichment of GAG contents than TGF-β3 at 7 days, but this trend was reversed at 14 days. Histological analysis results showed that TGF-β3 and 20B09 both allowed the efficient expression of proteoglycan by Safranin-O. Immunohistochemistry results showed that both collagenⅠ and collagenⅡ observed the high expression in 20B09, 20F08 treated groups, compared to similar expression levels in TGF-β3 treated groups. RT-PCR assay results supported the trend too. TGF-β3 groups showed a strong effect to chondrogenesis but transcripts for hypertrophy and osteogenesis were also increased. On the other hand, the selected small molecules 20B09, 20F08 did not. Therefore, small molecules can selectively induce the chondrogenic differentiation of hBM-MSCs. A detailed mechanistic research and target identification are currently under way and will be reported in due course. This experiment using small molecules is considered as the best substance to solve signaling problems on chondrogenesis.