CAJ | 학술논문

目的建立一种持续、稳定的可多向分化的骨髓间充质干细胞(BMSCs)体外分离培养体系.方法运用密度梯度离心法从1月龄新西兰兔骨髓中分离培养BMSCs,利用相差显微镜观察其形态及生长情况,扫描电镜观察其细胞结构,运用流式细胞术分析分离细胞群所处细胞周期和细胞活力,用MTT比色法绘制细胞生长曲线,并用特定诱导液将分离的BMSCs向成骨细胞和成脂肪细胞定向诱导分化,利用ALP和油红O进行染色鉴定.结果所分离的BMSCs细胞在形态学观察与生长动力学上均符合BMSCs特征,分离培养的BMSCs细胞在第3天进入对数生长期,第10天进入平台期;在成骨、成脂肪的诱导培养条件下,分别出现成骨、成脂肪表型特征,可进一步定向分化,结论所收获的细胞具有BMSCs的特异性.
목적 건립일충지속、은정적가다향분화적골수간충질간세포(BMSCs)체외분리배양체계.방법 운용밀도제도리심법종1월령신서란토골수중분리배양BMSCs,이용상차현미경관찰기형태급생장정황,소묘전경관찰기세포결구,운용류식세포술분석분리세포군소처세포주기화세포활력,용MTT비색법회제세포생장곡선,병용특정유도액장분리적BMSCs향성골세포화성지방세포정향유도분화,이용ALP화유홍O진행염색감정.결과 소분리적BMSCs세포재형태학관찰여생장동역학상균부합BMSCs특정,분리배양적BMSCs세포재제3천진입대수생장기,제10천진입평태기;재성골、성지방적유도배양조건하,분별출현성골、성지방표형특정,가진일보정향분화,결론소수획적세포구유BMSCs적특이성.
Objective To establish a sustained and stable bone marrow-derived mesenchymal stem cells (BMSCs) isolation and culture system which could be pluripotential in vitro. Methods BMSCs were isolated and cultured from the 1-month-old New Zealand white rabbits through density-gradient centrifugation. Growth and ultromicrostructure of the BMSCs were observed by light-microscope and scan electron-microscope. The cell activities and cell cycle were observed by flow cytometry. The grow curve of the BMSCs was described through MTT assay. BMSCs were induced and differentiate into osteoblasts and adipocytes by specific induction culture medium. Then the cells were stained with alkaline phosphatase (ALP) and Oil red O , and analyzed the result of all staining. Results The isolated BMSCs cells were in line with the characteristics of BMSCs in morphology and growth kinetics, and the isolated BMSCs cells would be into the logarithmic growth phase in the third day and into the platform phase in the tenth day. Conclusions In the osteogenic and adipogenic-induced culture conditions, BMSCs have appeared the phenotypic characteristics of osteogenic and adipogenic respectively, and could be further directed differentiation. These indicated the harvested cells possessed BMSCs specificity.