CAJ | 학술논문

背景：目前椎间盘髓核组织的细胞组成和特性仍未阐明。 　　目的：旨在建立大鼠椎间盘髓核来源间充质干细胞的体外培养体系，并对其体外多项分化潜能进行鉴定。 　　方法：体外培养SD大鼠盘髓核来源间充质干细胞，取第3代细胞进行三系诱导分化，将成骨、成脂和成软骨分化作为实验组，基础细胞培养作为对照组。 　　结果与结论：低密度培养获得的髓核来源细胞早期可形成葵花样细胞集落，克隆样生长。第3代后，细胞形态趋向均一，呈成纤维细胞样生长。成骨诱导28 d，实验组茜素红染色阳性，且 RunX2、osteopontin 及osteocalcin表达较对照组显著增高(P<0.05)；成脂诱导21 d，实验组油红O染色阳性，C/EBPα及PPARγ2表达较对照组显著增高(P <0.05)；成软骨分化诱导21 d，实验组番红O快绿染色和Ⅱ型胶原免疫组织化学染色阳性，aggrecan和Col2a1表达较对照组显著增高(P<0.05)。可见成年SD大鼠椎间盘髓核组织中可分离培养出体外呈克隆样生长的细胞群，且有向脂肪细胞、成骨细胞和软骨细胞发生定向分化的潜能。
배경：목전추간반수핵조직적세포조성화특성잉미천명。 　　목적：지재건립대서추간반수핵래원간충질간세포적체외배양체계，병대기체외다항분화잠능진행감정。 　　방법：체외배양SD대서반수핵래원간충질간세포，취제3대세포진행삼계유도분화，장성골、성지화성연골분화작위실험조，기출세포배양작위대조조。 　　결과여결론：저밀도배양획득적수핵래원세포조기가형성규화양세포집락，극륭양생장。제3대후，세포형태추향균일，정성섬유세포양생장。성골유도28 d，실험조천소홍염색양성，차 RunX2、osteopontin 급osteocalcin표체교대조조현저증고(P<0.05)；성지유도21 d，실험조유홍O염색양성，C/EBPα급PPARγ2표체교대조조현저증고(P <0.05)；성연골분화유도21 d，실험조번홍O쾌록염색화Ⅱ형효원면역조직화학염색양성，aggrecan화Col2a1표체교대조조현저증고(P<0.05)。가견성년SD대서추간반수핵조직중가분리배양출체외정극륭양생장적세포군，차유향지방세포、성골세포화연골세포발생정향분화적잠능。
BACKGROUND:Currently, cellular composition and the features of the nucleus pulposus are stil not to be clarified. OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential. METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls. RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic induction, safranin O/fast green staining was positive in the experimental group. The mRNA expressions of aggrecan and Col2a1 were significantly increased in the experimental group, compared to the control group (P<0.05). Our findings in this study suggested that nucleus pulposus-derived mesenchymal stem cells could be isolated from the Sprague-Dawley rat nucleus pulposus and exhibited clonal-like growth when they were cultured in vitro. These cells were confirmed to have the potential to differentiate into adipocytes, osteoblasts and chondrocytes in vitro.