CAJ | 학술논문

目的：建立小鼠骨片间充质干细胞(MSC)分离培养及扩增的方法。方法取小鼠胫骨和股骨，洗去骨髓后，用胶原酶Ⅰ消化疏松骨密质，利用MSC具有迁徙和贴壁生长的能力进行分离。并对获取的细胞进行流式鉴定和诱导分化。结果培养2d小鼠骨片边缘爬出成纤维样细胞，呈克隆和鱼群样生长，并可以进行持续传代培养。流式鉴定结果显示这群细胞表达MSC标志Scal1（92.7﹪），CD29（98.4﹪），CD90（91.6﹪），不表达造血细胞标志CD34（1.57﹪），CD45（3.99﹪），CD11b（0.63﹪），并可成功诱导分化成骨细胞和脂肪细胞。结论成功建立从小鼠骨片中获得MSC的方法，为实验研究提供可靠的细胞来源。
목적：건립소서골편간충질간세포(MSC)분리배양급확증적방법。방법취소서경골화고골，세거골수후，용효원매Ⅰ소화소송골밀질，이용MSC구유천사화첩벽생장적능력진행분리。병대획취적세포진행류식감정화유도분화。결과배양2d소서골편변연파출성섬유양세포，정극륭화어군양생장，병가이진행지속전대배양。류식감정결과현시저군세포표체MSC표지Scal1（92.7﹪），CD29（98.4﹪），CD90（91.6﹪），불표체조혈세포표지CD34（1.57﹪），CD45（3.99﹪），CD11b（0.63﹪），병가성공유도분화성골세포화지방세포。결론성공건립종소서골편중획득MSC적방법，위실험연구제공가고적세포래원。
Objective To establish a method for isolation and cultivation of mesenchymal stem cells from mouse bone. Methods Methods Tibia and femur were harvested from C57BL/6 mice, and bone marrow was flushed out of the cavity with α-MEM medium. The bone was cut into fragments with scissors and digested with collagenaseⅠfor 1-2 hours. The digested bone fragments were cultured in the medium for mesenchymal stem cells. The harvested cells were characterized by flow cytometry and differentiation capacity. Results Fibroblast-like cells migrated out of the bone fragments, which grew in the medium clonally and was cultured for several passages. Flow cytometry showed that the cells expressed Scal-1 (92.7 ﹪ ), CD29 (98.4﹪), CD90 (91.6﹪), but negative for hematopoietic cells markers CD34 (1.57﹪), CD45 (3.99 ﹪ ), CD11b (0.63 ﹪ ). And cells could be induced to differentiate into osteoblast and adipocyte. Conclusion Mesenchymal stem cells were successfully isolated from mouse bone fragments.