中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
7期
1339-1342
,共4页
段德宇%杨述华%吴星火%张志才%陈超%苏正兵%方为志
段德宇%楊述華%吳星火%張誌纔%陳超%囌正兵%方為誌
단덕우%양술화%오성화%장지재%진초%소정병%방위지
去分化重编排%成骨分化%脂肪干细胞%脐带静脉内皮细胞%共培养
去分化重編排%成骨分化%脂肪榦細胞%臍帶靜脈內皮細胞%共培養
거분화중편배%성골분화%지방간세포%제대정맥내피세포%공배양
Dedifferentiation-reprogrammed%Osteogenic differentiation%Adipose-derived stem cells%Umbilical vein endothelial cells%Co-culture
目的 观察脐带静脉内皮细胞(HUVEC)诱导去分化重编排脂肪干细胞成骨分化的效果并探讨其分子机制.方法 分离人脂肪干细胞(hASCs),取第3代hASCs进行去分化重编排并标记为去分化重编排脂肪干细胞(De-hASCs).分离人HUVEC,将hASCs与HUVEC以1∶1比例进行共培养(hASCs组),将De-hASCs与HUVEC以1∶1比例进行共培养(De-hASCs组).并于共培养4d和7d后检测碱性磷酸酶(ALP)活性,Runt相关基因2(Runx2)和骨形态发生蛋白-2(BMP-2)的mRNA表达水平,骨钙素的mRNA表达水平.结果 共培养4d后,De-hASCs组的ALP活性高于hASCs组,但差异无统计学意义(P＞0.05).共培养7d后,De-hASCs组ALP活性高于hASCs组,且差异有统计学意义(P＜0.05).共培养4d和7d后,De-hASCs组Runx2和BMP-2的mRNA表达水平均明显高于hASCs组,且差异有统计学意义(P＜0.05).共培养4d和7d后,De-hASCs组骨钙素的mRNA表达水平高于hASCs组,但差异无统计学意义(P＜0.05).结论 去分化重编排hASCs与HUVEC共培养条件下具有比普通脂肪干细胞更强的成骨分化能力,且与其上调BMP-2基因表达相关.
目的 觀察臍帶靜脈內皮細胞(HUVEC)誘導去分化重編排脂肪榦細胞成骨分化的效果併探討其分子機製.方法 分離人脂肪榦細胞(hASCs),取第3代hASCs進行去分化重編排併標記為去分化重編排脂肪榦細胞(De-hASCs).分離人HUVEC,將hASCs與HUVEC以1∶1比例進行共培養(hASCs組),將De-hASCs與HUVEC以1∶1比例進行共培養(De-hASCs組).併于共培養4d和7d後檢測堿性燐痠酶(ALP)活性,Runt相關基因2(Runx2)和骨形態髮生蛋白-2(BMP-2)的mRNA錶達水平,骨鈣素的mRNA錶達水平.結果 共培養4d後,De-hASCs組的ALP活性高于hASCs組,但差異無統計學意義(P＞0.05).共培養7d後,De-hASCs組ALP活性高于hASCs組,且差異有統計學意義(P＜0.05).共培養4d和7d後,De-hASCs組Runx2和BMP-2的mRNA錶達水平均明顯高于hASCs組,且差異有統計學意義(P＜0.05).共培養4d和7d後,De-hASCs組骨鈣素的mRNA錶達水平高于hASCs組,但差異無統計學意義(P＜0.05).結論 去分化重編排hASCs與HUVEC共培養條件下具有比普通脂肪榦細胞更彊的成骨分化能力,且與其上調BMP-2基因錶達相關.
목적 관찰제대정맥내피세포(HUVEC)유도거분화중편배지방간세포성골분화적효과병탐토기분자궤제.방법 분리인지방간세포(hASCs),취제3대hASCs진행거분화중편배병표기위거분화중편배지방간세포(De-hASCs).분리인HUVEC,장hASCs여HUVEC이1∶1비례진행공배양(hASCs조),장De-hASCs여HUVEC이1∶1비례진행공배양(De-hASCs조).병우공배양4d화7d후검측감성린산매(ALP)활성,Runt상관기인2(Runx2)화골형태발생단백-2(BMP-2)적mRNA표체수평,골개소적mRNA표체수평.결과 공배양4d후,De-hASCs조적ALP활성고우hASCs조,단차이무통계학의의(P＞0.05).공배양7d후,De-hASCs조ALP활성고우hASCs조,차차이유통계학의의(P＜0.05).공배양4d화7d후,De-hASCs조Runx2화BMP-2적mRNA표체수평균명현고우hASCs조,차차이유통계학의의(P＜0.05).공배양4d화7d후,De-hASCs조골개소적mRNA표체수평고우hASCs조,단차이무통계학의의(P＜0.05).결론 거분화중편배hASCs여HUVEC공배양조건하구유비보통지방간세포경강적성골분화능력,차여기상조BMP-2기인표체상관.
Objective To study the osteogenic differentiation and related molecular mechanisms of dedifferentiation reprogrammed hASCs (De-hASCs) induced by human umbilical vein endothelial cells (HUVECs).Methods HASCs were isolated from 6 volunteers.HASCs passaged three times were then selected to undergo the dedifferentiation reprogrammed process.These cells were denoted as the De-hASCs.Both De-hASCs and hASCs were then co-cultured with HUVECs for a total period of one week.Alkaline phosphatase (ALP) activity of cells cultured for 4 and 7 days were measured.runt related transcription factor-2 (Runx2) and bone morphogenetic protein-2 (BMP-2) mRNA levels of cells cultured for 4 and 7 days were detected.Osteocalcin mRNA expression levels of cells cultured for 4 and 7 days were examined.Results ALP activity of the De-hASCs measured 4 days after cell culture was stronger than that of hASCs,but there was no significant difference.ALP acitivty of the De-hASCs measured 7 days after cell culture was stronger than that of hASCs with significant difference.Runx2 and BMP-2 mRNA expression levels of the De-hASCs were higher than those of hASCs both 4 and 7 days after cell culture with siginificant difference.Osteocalcin mRNA expression levels of De-hASCs were higher than those of hASCs both 4 and 7 days after cell culture but without significant difference.Conclusion When co-cultured with HUVECs,De-hASCs possessed a stronger osteogenic differentiation capacity than hASCs by up-regulating the expression of BMP-2.
