中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2010年
8期
752-756
,共5页
高杰%韩建伟%朱洪繁%杨彤涛%范清宇%马保安
高傑%韓建偉%硃洪繁%楊彤濤%範清宇%馬保安
고걸%한건위%주홍번%양동도%범청우%마보안
骨髓%间充质干细胞%microRNA
骨髓%間充質榦細胞%microRNA
골수%간충질간세포%microRNA
Bone marrow%Mesenchymal stem cells%MicroRNA
目的 筛选可作为人骨髓间充质干细胞(MSCs)分子标记物的microRNA(miRNA).方法 以从骨髓中分离培养的MSCs及其诱导分化的成骨细胞和软骨细胞为实验对象,利用基因芯片检测miRNA的表达情况,由芯片显著性分析(SAM)得到MSCs较其诱导分化细胞高表达的miRNA,再由实时定量PCR进行验证.结果 成功分离培养出MSCs,并分别诱导其分化为成骨细胞和软骨细胞;MSCs及由其诱导分化的成骨细胞和软骨细胞经miRNA芯片检测及SAM分析得到8个MSCs较由其诱导分化的成骨细胞高表达miRNA(has-miR-424、has-miR-34a、has-miR-593、has-miR-10a、has-miR-148a、has-miR-602、mmu-miR-709、mmu-miR-665),3个MSCs较由其诱导分化的软骨细胞高表达miRNA(has-miR-424、PREDICTED_MIR189、mmu-miR-665).其中MSCs较成骨细胞和软骨细胞均高表达的人源性miRNA为has-miR-424,差异倍数分别为6.6倍和4.4倍;在原样本中对进行实时定量PCR的验证,结果提示与诱导分化的成骨细胞相比,MSCs中的has-miR-424表达升高约3.6倍,而与诱导分化的软骨细胞相比,MSCs中的has-miR-424表达升高约3.1倍,结果与芯片结果相符合.结论 MSCs较其诱导分化细胞高表达的miRNA有望成为人骨髓间充质干细胞的一种特异性的分子标记物,这些miRNA对MSCs自我更新和未分化状态起着重要的维持作用.
目的 篩選可作為人骨髓間充質榦細胞(MSCs)分子標記物的microRNA(miRNA).方法 以從骨髓中分離培養的MSCs及其誘導分化的成骨細胞和軟骨細胞為實驗對象,利用基因芯片檢測miRNA的錶達情況,由芯片顯著性分析(SAM)得到MSCs較其誘導分化細胞高錶達的miRNA,再由實時定量PCR進行驗證.結果 成功分離培養齣MSCs,併分彆誘導其分化為成骨細胞和軟骨細胞;MSCs及由其誘導分化的成骨細胞和軟骨細胞經miRNA芯片檢測及SAM分析得到8箇MSCs較由其誘導分化的成骨細胞高錶達miRNA(has-miR-424、has-miR-34a、has-miR-593、has-miR-10a、has-miR-148a、has-miR-602、mmu-miR-709、mmu-miR-665),3箇MSCs較由其誘導分化的軟骨細胞高錶達miRNA(has-miR-424、PREDICTED_MIR189、mmu-miR-665).其中MSCs較成骨細胞和軟骨細胞均高錶達的人源性miRNA為has-miR-424,差異倍數分彆為6.6倍和4.4倍;在原樣本中對進行實時定量PCR的驗證,結果提示與誘導分化的成骨細胞相比,MSCs中的has-miR-424錶達升高約3.6倍,而與誘導分化的軟骨細胞相比,MSCs中的has-miR-424錶達升高約3.1倍,結果與芯片結果相符閤.結論 MSCs較其誘導分化細胞高錶達的miRNA有望成為人骨髓間充質榦細胞的一種特異性的分子標記物,這些miRNA對MSCs自我更新和未分化狀態起著重要的維持作用.
목적 사선가작위인골수간충질간세포(MSCs)분자표기물적microRNA(miRNA).방법 이종골수중분리배양적MSCs급기유도분화적성골세포화연골세포위실험대상,이용기인심편검측miRNA적표체정황,유심편현저성분석(SAM)득도MSCs교기유도분화세포고표체적miRNA,재유실시정량PCR진행험증.결과 성공분리배양출MSCs,병분별유도기분화위성골세포화연골세포;MSCs급유기유도분화적성골세포화연골세포경miRNA심편검측급SAM분석득도8개MSCs교유기유도분화적성골세포고표체miRNA(has-miR-424、has-miR-34a、has-miR-593、has-miR-10a、has-miR-148a、has-miR-602、mmu-miR-709、mmu-miR-665),3개MSCs교유기유도분화적연골세포고표체miRNA(has-miR-424、PREDICTED_MIR189、mmu-miR-665).기중MSCs교성골세포화연골세포균고표체적인원성miRNA위has-miR-424,차이배수분별위6.6배화4.4배;재원양본중대진행실시정량PCR적험증,결과제시여유도분화적성골세포상비,MSCs중적has-miR-424표체승고약3.6배,이여유도분화적연골세포상비,MSCs중적has-miR-424표체승고약3.1배,결과여심편결과상부합.결론 MSCs교기유도분화세포고표체적miRNA유망성위인골수간충질간세포적일충특이성적분자표기물,저사miRNA대MSCs자아경신화미분화상태기착중요적유지작용.
Objective To identify the specific microRNA (miRNA) that can be taken as a molecular marker for human bone marrow-derived mesenchymal stem cells (MSCs). Methods MSCs were isolated and cultured from bone marrow through density centrifugation and then were induced to differentiate into osteoblasts and chondrocytes. Samples of MSCs, osteoblasts and chondrocytes were detected by miRNA microarrays single channel fluorescence chip to determine the expression levels of miRNAs. Significance Analysis of Microarrays ( SAM, version 2.1 ) software was used to analyze the raw data to determine the miRNAs overexpressed in MSCs, which was validated in the same sample using real time reserve transcriptase polymerase chain reaction (RT-PCR). Results MSCs were successfully isolated from bone marrow and induced to differentiate into osteoblasts and chondrocytes in vitro. Microarrays showed that eight miRNAs (has-miR-424, has-miR-34a, has-miR-593, has-miR-10a, has-miR-148a,has-miR-602, mmu-miR-709 and mmu-miR-665) were overexpressed in MSCs but underexpressed in osteoblasts. Three miRNAs including has-miR-424, PREDICTED_MIR189 and mmu-miR-665, were overexpressed in MSCs but underexpressed in chondrocytes. The has-miR-424 expression in MSCs was 6.6times higher than in osteoblasts and 4.4 times higher than in chondrocytes. The results of real time RTPCR showed that the miR-424 was overexpressed in MSCs, 3.6 times higher than that in osteoblasts and 3.1 times higher than that in chondrocytes, which was coincident with the results of microarray. Conclusions The screened MSCs express more miRNAs in comparison with osteoblasts and chondrocytes,play important roles in maintaining self renewal and undifferentiation of MSCs and is a promising specific molecule marker for MSCs.
