目的比较F icoll(1.073 g/m l)密度梯度离心法分离人骨髓间充质干细胞(hum an bone m arrow derived m esen-chym al stem cells,hBMMSCs)与常用的Percoll分离法之间的差异。建立一种简便、实用的hBMMSCs分离方法。方法分别应用上述两...目的比较F icoll(1.073 g/m l)密度梯度离心法分离人骨髓间充质干细胞(hum an bone m arrow derived m esen-chym al stem cells,hBMMSCs)与常用的Percoll分离法之间的差异。建立一种简便、实用的hBMMSCs分离方法。方法分别应用上述两种方法分离hBMMSCs,比较用两种方法从骨髓中分离的有核细胞得率及死亡率、贴壁细胞克隆数、细胞形态以及细胞表面标志。结果两种方法获得的有核细胞得率无显著差异(P>0.05);F icoll(1.073 g/m l)密度梯度离心法获得的有核细胞死亡率显著小于Percoll分离法(P<0.01);F icoll(1.073 g/m l)密度梯度离心法获得的hBMMSCs原代培养5 d的贴壁细胞克隆数/37.5 cm2显著多于Percoll分离法(P<0.05)。F icoll(1.073 g/m l)密度梯度离心法与Percoll分离法获得的hBMMSCs细胞形态均一,为纺锤形或三角形;F icoll(1.073 g/m l)密度梯度离心法与Percoll分离法获得的原代hBMMSCs CD105阳性表达率[(94.0±2.0)%vs(95.8±1.5)%]及CD34阴性表达率[(96.0±1.2)%vs(97±1.0)%]无显著差异(P>0.05)。结论与Percoll分离法相比较,F icoll(1.073 g/m l)密度梯度离心法分离hBMMSCs具有细胞死亡率较小,细胞得率较多的优点,是一种较好的hBMMSCs分离方法。展开更多
On a woman with severe intrauterine adhesions, hysteroscopy followed by cyclical hormone replacement therapy was tried for 5 months, for development of the endometrium. When this failed, autologous stem cells were tri...On a woman with severe intrauterine adhesions, hysteroscopy followed by cyclical hormone replacement therapy was tried for 5 months, for development of the endometrium. When this failed, autologous stem cells were tried as an alternative therapy. Adult autologous bone marrow mesenchymal stem cells isolated from patient’s own bone marrow and were cultured and placed in the endometrial cavity under ultrasound guidance after curettage. Patient was then given cyclical hormonal therapy. Endometrium was assessed intermittently using ultrasound. Three months later, endometrium partly recovered with improved ultrasonic echo. This resulted in spontaneous pregnancy followed by confirmation of gestational sac, yolk sac, and primitive heart tube pulse on ultrasound. Autologous bone marrow derived mesenchymal stem cells could regenerate injured endometrium not responding to conventional treatment and can be used as an alternative in females with severe Asherman’s syndrome.展开更多
目的:原代培养并鉴定大鼠骨髓间充质干细胞作为骨组织工程的种子细胞。方法:取大鼠骨髓,贴壁培养法培养。绘制生长曲线,检测细胞周期。real time PCR检测不同代数BMSCs胞内成骨相关因子mRNA水平。流式细胞仪检测细胞表面标志物。并行成...目的:原代培养并鉴定大鼠骨髓间充质干细胞作为骨组织工程的种子细胞。方法:取大鼠骨髓,贴壁培养法培养。绘制生长曲线,检测细胞周期。real time PCR检测不同代数BMSCs胞内成骨相关因子mRNA水平。流式细胞仪检测细胞表面标志物。并行成脂及成骨诱导。结果:原代取材后24 h细胞即可贴壁,7 d可生长达培养皿底面积的80%。P3代BMSCs的生长曲线呈S形,近90%的细胞处于G0/G1期。从P0到P5BMSCs中几乎都有内源性bFGF、Shh、Cbfa1、ALP、OC的表达。流式细胞仪检测99.7%的BMSCs胞膜表面同时表达CD29及CD90,不表达CD45。并具有成骨分化及成脂分化的能力。结论:原代培养的BMSCs为骨组织工程提供了可靠的细胞来源。展开更多
文摘On a woman with severe intrauterine adhesions, hysteroscopy followed by cyclical hormone replacement therapy was tried for 5 months, for development of the endometrium. When this failed, autologous stem cells were tried as an alternative therapy. Adult autologous bone marrow mesenchymal stem cells isolated from patient’s own bone marrow and were cultured and placed in the endometrial cavity under ultrasound guidance after curettage. Patient was then given cyclical hormonal therapy. Endometrium was assessed intermittently using ultrasound. Three months later, endometrium partly recovered with improved ultrasonic echo. This resulted in spontaneous pregnancy followed by confirmation of gestational sac, yolk sac, and primitive heart tube pulse on ultrasound. Autologous bone marrow derived mesenchymal stem cells could regenerate injured endometrium not responding to conventional treatment and can be used as an alternative in females with severe Asherman’s syndrome.
文摘目的:原代培养并鉴定大鼠骨髓间充质干细胞作为骨组织工程的种子细胞。方法:取大鼠骨髓,贴壁培养法培养。绘制生长曲线,检测细胞周期。real time PCR检测不同代数BMSCs胞内成骨相关因子mRNA水平。流式细胞仪检测细胞表面标志物。并行成脂及成骨诱导。结果:原代取材后24 h细胞即可贴壁,7 d可生长达培养皿底面积的80%。P3代BMSCs的生长曲线呈S形,近90%的细胞处于G0/G1期。从P0到P5BMSCs中几乎都有内源性bFGF、Shh、Cbfa1、ALP、OC的表达。流式细胞仪检测99.7%的BMSCs胞膜表面同时表达CD29及CD90,不表达CD45。并具有成骨分化及成脂分化的能力。结论:原代培养的BMSCs为骨组织工程提供了可靠的细胞来源。
