인간간엽줄기세포의 연골세포 분화 유도 성장인자 및 주사침 크기 차이에 따른 세포반응에 대한 in vitro 연구
A study of growth factors, chondrogenic differentiation of mesenchymal stem cells and cell response by needle size differences in vitro
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"목적: 인간간엽줄기세포를 연골세포로 분화유도하는 성장인자 규명 및 주사침 크기 차이에 따른 세포증식 증진 비교이다. 연구 재료 및 방법: 인간간엽줄기세포를 연골세포유도배지에서 14, 21, 28일 배양하여 BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1. TGF-β2, TGF-β3를 투여한 군들과 대조군에서 SOX-5, SOX-6, SOX-9 및 FOXO1A의 발현량을 분석하였다. 18, 21, 26 게이지(G) 주사침을 사용하여 배양접시에 인간간엽줄기세포를 접종하고 24, 48, 72시간 후 각각 세포증식을 측정하였다. 결과: 기존에 알려진 FGF, IGF-1, TGFβ1와 함께, BMP-2, BMP-4, BMP-6, BMP-7 등 BMP family 성장인자들에 의하여 연골세포분화 전사인자 유전자들인 SOX-5, SOX-6, SOX-9, FOXO1A의 유전자발현이 증가하였다. 48시간에서는 가장 작은 주사침인 26G군이 대조군 및 18G군에 비하여 유의한 세포증식 증진을 보였으며, 72시간에서도 가장 작은 주사침인 26G군이 대조군에 비하여 유의한 세포증식 증진을 보였다. 결론: 인간간엽줄기세포의 연골세포분화 유도능을 가진 성장인자들을 고찰할 수 있었고 주사침 크기에 따른 세포증식 변화를 규명할 수 있었다."
Abstract
Purpose: This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences. Materials and Methods: Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively. Results: In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell roliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group. Conclusion: Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.
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