CAJ | 학술논문

目的：探讨短期冻存对兔骨髓间充质干细胞（BMSCs）的增殖、生物特征及成骨能力的影响。方法选取第5代兔BMSCs冻存，30 d后复苏、体外培养至第10代作为实验组，设相同代数未冻存的BMSCs为对照组。MTT、细胞周期、细胞表面标志及成骨诱导分化后碱性磷酸酶（ALP）水平、Ⅰ型胶原及钙结节染色作为其增殖、生物特征及成骨能力的评价指标。结果冻存后BM SCs生长潜伏期延长，但生长状态无明显影响；增殖指数和G1期细胞所占比例分别为42．9％±3．4％、57．0％±3．4％，细胞表面标志CD44阳性率为93．62％±1．05％，不表达CD45；成骨诱导第7、14天 ALP水平（U/gprot）分别为6．73±1．92、15．99±4．36，第14、21天，Ⅰ型胶原、钙结节茜素红染色阳性；以上结果与对照组比较差异均无统计学意义（ P＞0．05）。结论短期冻存对兔BM SCs增殖、生物特征及成骨分化潜能无明显影响，冻存后可继续进一步研究。
목적：탐토단기동존대토골수간충질간세포（BMSCs）적증식、생물특정급성골능력적영향。방법선취제5대토BMSCs동존，30 d후복소、체외배양지제10대작위실험조，설상동대수미동존적BMSCs위대조조。MTT、세포주기、세포표면표지급성골유도분화후감성린산매（ALP）수평、Ⅰ형효원급개결절염색작위기증식、생물특정급성골능력적평개지표。결과동존후BM SCs생장잠복기연장，단생장상태무명현영향；증식지수화G1기세포소점비례분별위42．9％±3．4％、57．0％±3．4％，세포표면표지CD44양성솔위93．62％±1．05％，불표체CD45；성골유도제7、14천 ALP수평（U/gprot）분별위6．73±1．92、15．99±4．36，제14、21천，Ⅰ형효원、개결절천소홍염색양성；이상결과여대조조비교차이균무통계학의의（ P＞0．05）。결론단기동존대토BM SCs증식、생물특정급성골분화잠능무명현영향，동존후가계속진일보연구。
Objective To study the effects of short-term cryopreservation on the proliferation ,bio-characteristics and osteogenic capability of rabbit BMSCs and lay the foundation for the further study .Methods The 5th generation of rabbit BMSCs were cryo-preservated by liquid nitrogen for 30 days ,and then thawing the cells to culture it to the 10th generation in vitro ,put the non-cryo-preservated and the same generation BMSCs as the control group .The MTT ,cell cycle ,cell surface marker ,the content of ALP and the immunohistochemical staining for collagen typeⅠ and alizarin red staining for calcium after differentiation induced by osteogene-sis were used to evaluate the proliferation ,bio-characteristic and osteogenic differentiation capability of rabbit BMSCs .Results The growth incubation period of BMSCs after cryopreservated was extended ,but it gradually recover through serial passage .The prolif-eration index and the proportion of BMSCs in G1 phase were 42 .9% ± 3 .4% and 57 .0% ± 3 .4% respectively .The positive rate of cell surface marker of CD44 was 93 .62% ± 1 .05% without expression of CD45 .The contents of ALP(U/gprot) were 6 .73 ± 1 .92 and 15 .99 ± 4 .36 in BMSCs after 7th day and 14th day with osteogenic induction ,respectively .The collagen typeⅠ and alizarin red staining for calcium indicated positive at BMSCs after 14th day and 21th day with osteogenic induction ,respectively .These results showed no significant difference compared with the control group (P>0 .05) .Conclusion Short-term cryopreservation has no obvi-ous impacts on the proliferation ,bio-characteristic and osteogenic capability of BMSCs and it could be used for further study .