中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
9期
1983-1985
,共3页
转化生长因子-β2%转化生长因子-β1%骨髓间充质干细胞%成骨细胞
轉化生長因子-β2%轉化生長因子-β1%骨髓間充質榦細胞%成骨細胞
전화생장인자-β2%전화생장인자-β1%골수간충질간세포%성골세포
Transforming growth factor-β2%Transforming growth factor-β1%Bone marrow mesenchymal stem cells%Osteoblast
目的 观察转化生长因子(TGF)-β2在体外诱导骨髓间充质干细胞(BMSCs)向成骨细胞分化能力.方法 取第3代BMSCs接种于3.5×103个/孔密度培养板,分为3组:A、B、C组,A组不做任何处理,B、C组培养基中分别添加5 μg/L TGF-β1、5μg/L TGF-β2,每孔200μl,每组4孔.7d对3个实验组行碱性磷酸酶(ALP)活性检测和Western blot检测Collagen Ⅰ表达,第3代BMSCs以5×107/L细胞浓度接种于6孔培养板,每组1板,分组添加方式同前,21d细胞爬片行Von kossa染色.结果 A、B、C组第7天的ALP活力值分别为1.15±0.16、4.26±0.39、4.34±0.14,各组间比较差异有统计学意义(P＜0.05),ALP的染色面积及深度、Von kossa染色钙化结节C组强于B组和A组,B组强于A组;Collagen Ⅰ表达C组高于B组和A组,B组高于A组(P＜0.05).结论 在一定条件下,TGF-β2体外诱导BMSCs成骨能力较转化生长因子-β1强.
目的 觀察轉化生長因子(TGF)-β2在體外誘導骨髓間充質榦細胞(BMSCs)嚮成骨細胞分化能力.方法 取第3代BMSCs接種于3.5×103箇/孔密度培養闆,分為3組:A、B、C組,A組不做任何處理,B、C組培養基中分彆添加5 μg/L TGF-β1、5μg/L TGF-β2,每孔200μl,每組4孔.7d對3箇實驗組行堿性燐痠酶(ALP)活性檢測和Western blot檢測Collagen Ⅰ錶達,第3代BMSCs以5×107/L細胞濃度接種于6孔培養闆,每組1闆,分組添加方式同前,21d細胞爬片行Von kossa染色.結果 A、B、C組第7天的ALP活力值分彆為1.15±0.16、4.26±0.39、4.34±0.14,各組間比較差異有統計學意義(P＜0.05),ALP的染色麵積及深度、Von kossa染色鈣化結節C組彊于B組和A組,B組彊于A組;Collagen Ⅰ錶達C組高于B組和A組,B組高于A組(P＜0.05).結論 在一定條件下,TGF-β2體外誘導BMSCs成骨能力較轉化生長因子-β1彊.
목적 관찰전화생장인자(TGF)-β2재체외유도골수간충질간세포(BMSCs)향성골세포분화능력.방법 취제3대BMSCs접충우3.5×103개/공밀도배양판,분위3조:A、B、C조,A조불주임하처리,B、C조배양기중분별첨가5 μg/L TGF-β1、5μg/L TGF-β2,매공200μl,매조4공.7d대3개실험조행감성린산매(ALP)활성검측화Western blot검측Collagen Ⅰ표체,제3대BMSCs이5×107/L세포농도접충우6공배양판,매조1판,분조첨가방식동전,21d세포파편행Von kossa염색.결과 A、B、C조제7천적ALP활력치분별위1.15±0.16、4.26±0.39、4.34±0.14,각조간비교차이유통계학의의(P＜0.05),ALP적염색면적급심도、Von kossa염색개화결절C조강우B조화A조,B조강우A조;Collagen Ⅰ표체C조고우B조화A조,B조고우A조(P＜0.05).결론 재일정조건하,TGF-β2체외유도BMSCs성골능력교전화생장인자-β1강.
Objective To investigate the ability of transforming growth factor (TGF)-β2 to differentiate the bone marrow mesenchymal stem cells (BMSCs) into bone cells in vitro.Methods BMSCs of 3rd generation were seeded in 3.5 × 103/pore density culture plate and divided into three groups:A,B and C.Group A received no treatment.Groups B and C received the culture medium containing 5 μg/L TGF-β1 and 5 μg/L TGF-β2 respectively,200 μL per well of each of the four holes.On 7th day,alkaline phosphatase (ALP) activity assay and Western blotting method were applied to detect the expression of collagen Ⅰ.BMSCs of 3rd generation were seeded in six well plates at a density of 5 × 107/L,grouping and treatment principle were performed as described before,and the calcium salts were observed by Von Kassa at the 21st day.Results The ALP activity values in groups A,B and C on 7th day were 1.15 ± 0.16,4.26 ± 0.39 and 4.34 ± 0.14 respectively (P ＜ 0.05).The stained area and depth of ALP and Von kossa staining for calcified nodules were increased in group C as compared with groups A and B,and those in group B were increased as compared with group A.Collagen Ⅰ expression in group C was highest among the groups and lowest in group A.Conclusion Under above mentioned conditions,TGF-β2 induced BMSCs osteogenic capacity significantly stronger than TGF-β1 and the control.
