CAJ | 학술논문

背景：研究显示兔脂肪干细胞的体外分离方法大多数为Ⅰ型胶原酶消化法，采用组织块贴壁法扩增兔脂肪干细胞尚不多见。 　　目的：采用组织块贴壁法从兔脂肪组织中分离培养兔脂肪干细胞，并进行成脂、成骨的诱导分化。 　　方法：采用组织块贴壁法分离出兔脂肪干细胞，进行体外培养，观察其形态特征。取对数生长期的第3代细胞，用MTT法绘制其生长曲线；流式细胞仪检测其表面抗原CD29、CD44、CD45的表达情况；分别用成脂和成骨诱导培养液诱导其向脂肪细胞和成骨细胞分化，油红O、茜素红染色定性鉴定。 　　结果与结论：体外培养的兔脂肪干细胞呈梭形纤维样细胞形态，生长力旺盛。流式细胞术分析结果显示，第3代兔脂肪间充质干细胞强表达CD29，CD44，阴性表达CD45。兔脂肪干细胞经成脂诱导14 d后，油红O染色阳性；成骨诱导培养14 d后，茜素红染色阳性。提示组织块贴壁法可以分离培养出兔脂肪干细胞。
배경：연구현시토지방간세포적체외분리방법대다수위Ⅰ형효원매소화법，채용조직괴첩벽법확증토지방간세포상불다견。 　　목적：채용조직괴첩벽법종토지방조직중분리배양토지방간세포，병진행성지、성골적유도분화。 　　방법：채용조직괴첩벽법분리출토지방간세포，진행체외배양，관찰기형태특정。취대수생장기적제3대세포，용MTT법회제기생장곡선；류식세포의검측기표면항원CD29、CD44、CD45적표체정황；분별용성지화성골유도배양액유도기향지방세포화성골세포분화，유홍O、천소홍염색정성감정。 　　결과여결론：체외배양적토지방간세포정사형섬유양세포형태，생장력왕성。류식세포술분석결과현시，제3대토지방간충질간세포강표체CD29，CD44，음성표체CD45。토지방간세포경성지유도14 d후，유홍O염색양성；성골유도배양14 d후，천소홍염색양성。제시조직괴첩벽법가이분리배양출토지방간세포。
BACKGROUND:The rabbit adipose-derived stem cells are mostly isolated by type I col agenase digestion, but rarely by explants culture method. OBJECTIVE:To isolate rabbit adipose-derived stem cells for adipogenic and osteogenic differentiation. METHODS:The rabbit adipose-derived stem cells were isolated from rabbit adipose by explants culture method, and cultured in vitro fol owed by morphological observation. The grow curve and cellsurface markers CD29, CD44, CD45 of passage 3 cells were analyzed respectively by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and flow cytometry;cells from the third passages were induced for adipogenic and osteogenic differentiation by different revulsants, and cells were examined by oil red O staining and alizarin red staining . RESULTS AND CONCLUSION:The rabbit adipose-derived stem cells cultured in vitro exhibited a spindle-shaped appearance and could rapidly expand. Flow cytometry analysis revealed that the third passage of rabbit adipose-derived stem cells was positive for CD29, CD44, but negative for CD45. Rabbit adipose-derived stem cells were positive for oil red O staining at 14 days of adipogenic induction, and positive for alizarin red staining at 14 days of osteogenic induction. In conclusion, we could successful y isolate rabbit adipose-derived stem cells using explants culture method.