牡丹江医学院学报
牡丹江醫學院學報
모단강의학원학보
JOURNAL OF MUDANJIANG MEDICAL COLLEGE
2014年
4期
8-12
,共5页
脂肪来源干细胞%软骨细胞%共培养%软骨分化
脂肪來源榦細胞%軟骨細胞%共培養%軟骨分化
지방래원간세포%연골세포%공배양%연골분화
Adipose-derived stem cells%Chondrocytes%Co-culture%Chondrogenic differentiation
目的：验证脂肪来源干细胞（adipose-derived stem cells，ADSCs）在软骨细胞共培养后，能否诱导ADSCs向软骨细胞分化，并在此基础上形成软骨复合体和移植界面。方法：分离并培养成年健康新西兰大白兔的ADSCs和软骨细胞，实验分为两组，将ADSCs和软骨细胞共同培养作为实验组，单独培养ADSCs作为对照组。培养14d后，倒置相差显微镜观察两组ADSCs的形态变化；通过甲苯胺蓝染色法和免疫组织化学染色法进行成软骨诱导鉴定；实时荧光定量PCR法检测两组ADSCs蛋白多糖和Ⅱ型胶原的mRNA表达变化。结果：倒置相差显微镜观察显示，共培养组第14d时，细胞大部分呈圆形，呈软骨细胞形态；对照组ADSCs的形态未见明显变化仍以梭形为主，排列有序，呈束状或漩涡状排列，增殖迅速；成软骨鉴定结果显示，共培养组甲苯胺蓝染色、Ⅱ型胶原免疫组织化学染色均为阳性，而对照组均为阴性；实时荧光定量PCR检测显示，共培养组ADSCs蛋白多糖和Ⅱ型胶原的mRNA相对表达量均明显高于对照组。结论：与软骨细胞共同培养后，ADSCs可被诱导向软骨细胞分化，并在此基础上形成软骨复合体和移植界面。
目的：驗證脂肪來源榦細胞（adipose-derived stem cells，ADSCs）在軟骨細胞共培養後，能否誘導ADSCs嚮軟骨細胞分化，併在此基礎上形成軟骨複閤體和移植界麵。方法：分離併培養成年健康新西蘭大白兔的ADSCs和軟骨細胞，實驗分為兩組，將ADSCs和軟骨細胞共同培養作為實驗組，單獨培養ADSCs作為對照組。培養14d後，倒置相差顯微鏡觀察兩組ADSCs的形態變化；通過甲苯胺藍染色法和免疫組織化學染色法進行成軟骨誘導鑒定；實時熒光定量PCR法檢測兩組ADSCs蛋白多糖和Ⅱ型膠原的mRNA錶達變化。結果：倒置相差顯微鏡觀察顯示，共培養組第14d時，細胞大部分呈圓形，呈軟骨細胞形態；對照組ADSCs的形態未見明顯變化仍以梭形為主，排列有序，呈束狀或漩渦狀排列，增殖迅速；成軟骨鑒定結果顯示，共培養組甲苯胺藍染色、Ⅱ型膠原免疫組織化學染色均為暘性，而對照組均為陰性；實時熒光定量PCR檢測顯示，共培養組ADSCs蛋白多糖和Ⅱ型膠原的mRNA相對錶達量均明顯高于對照組。結論：與軟骨細胞共同培養後，ADSCs可被誘導嚮軟骨細胞分化，併在此基礎上形成軟骨複閤體和移植界麵。
목적：험증지방래원간세포（adipose-derived stem cells，ADSCs）재연골세포공배양후，능부유도ADSCs향연골세포분화，병재차기출상형성연골복합체화이식계면。방법：분리병배양성년건강신서란대백토적ADSCs화연골세포，실험분위량조，장ADSCs화연골세포공동배양작위실험조，단독배양ADSCs작위대조조。배양14d후，도치상차현미경관찰량조ADSCs적형태변화；통과갑분알람염색법화면역조직화학염색법진행성연골유도감정；실시형광정량PCR법검측량조ADSCs단백다당화Ⅱ형효원적mRNA표체변화。결과：도치상차현미경관찰현시，공배양조제14d시，세포대부분정원형，정연골세포형태；대조조ADSCs적형태미견명현변화잉이사형위주，배렬유서，정속상혹선와상배렬，증식신속；성연골감정결과현시，공배양조갑분알람염색、Ⅱ형효원면역조직화학염색균위양성，이대조조균위음성；실시형광정량PCR검측현시，공배양조ADSCs단백다당화Ⅱ형효원적mRNA상대표체량균명현고우대조조。결론：여연골세포공동배양후，ADSCs가피유도향연골세포분화，병재차기출상형성연골복합체화이식계면。
Objective:This study is to verify the chondrogenic differentiation of adipose -derived stem cells (ADSCs) by co-cul-turing chondrocytes and ADSCs .Methods:ADSCs and chondrocytes were isolated and cultured from adult healthy New Zealand rabbits . The experiment is divided into two groups .Chondrocytes and ADSCs were co -cultured in Transwell 6-well plates as the experimental group and ADSCs were cultured alone as the control group .The morphology changes of the ADSCs of the two groups were observed with inverted phase contrast microscope .Toluidine blue staining and immunohistochemistry staining were used to identify the chondrogenic differentiation of ADSCs , and the mRNA expressions of aggrecan and collagen type II were detected with real -time luorescent quanti-tative PCR.Results:By Day 14 after co-culturing, most ADSCs in the experimental group gradually manifested circular shape like cartilage cell in morphology;while ADSCs in the control group did not change obviously and most of them were fusiform , arranged or-derly, beam shaped or vortex pattern in the morphology with rapid proliferation .Cartilage identification results show that the results of toluidine blue staining and collagen type Ⅱ immunohistochemical staining were positive in the experimental group , while the control group were negative;Real-time fluorescent quantitative PCR detection showed that the mRNA expression levels of proteoglycan and collagen type Ⅱwere significantly higher than that in the control group .Conclusion:After co-culture with chondrocytes , ADSCs can be induced to differentiate into chondrocytes and form cartilage complex and transplantation interface based on them .
