Mesenchymal stem cells （MSCs） are multipotent stem cells capable of differentiating into various cell types,including osteocytes,chondrocytes,adipocytes,myocytes,and tenocytes.However,the difficulty or failure in ex...Mesenchymal stem cells （MSCs） are multipotent stem cells capable of differentiating into various cell types,including osteocytes,chondrocytes,adipocytes,myocytes,and tenocytes.However,the difficulty or failure in expanding the mouse MSCs in vitro greatly hampered important research in animal models.The OP9,a stromal cell line from mouse bone marrow,has hematopoietic supportive capacity.Here,we report that the OP9 has the immunophenotype （CD45-,CD11b-,FLK-1-,CD31-,CD34-,CD44＋,CD29＋,Sca-1＋,CD86-,and MHCII-） identical to canonical mouse MSCs.The expression of CD140a＋,CD140b＋,α-SMA＋ and Calponin＋ suggested the perivascular origin of OP9.Functionally,the OP9 had strong clonogenic ability and could be induced into osteocytes,chondrocytes and adipocytes.The lymphocyte transformation test （LTT） and mixed leukocyte reaction （MLR） showed that the OP9 could suppress T lymphocyte proliferation stimulated by nonspecific mitogens （PHA） or allogeneic lymphocytes （BALB/c T cells）.Finally,the migration of OP9 could be efficiently induced by bFGF,IGF-1,IL-3,PDGF-BB,TGF-β1 and TGF-β3.In conclusion,the OP9 were bona fide MSCs,and such homogenous cell line will be helpful to delineate biological features of MSCs at the stem cell level.展开更多
目的探讨表皮生长因子(EGF)对5-氟尿嘧啶(5-FU)化疗损伤小鼠骨髓基质细胞(OP9)的保护作用。方法首先建立5-FU化疗损伤OP9细胞模型并摸索EGF作用剂量。然后,将实验分为空白对照组、模型组、实验组。用25μg/m L 5-FU作用OP9细胞作为模型...目的探讨表皮生长因子(EGF)对5-氟尿嘧啶(5-FU)化疗损伤小鼠骨髓基质细胞(OP9)的保护作用。方法首先建立5-FU化疗损伤OP9细胞模型并摸索EGF作用剂量。然后,将实验分为空白对照组、模型组、实验组。用25μg/m L 5-FU作用OP9细胞作为模型组,用20 ng/m L的EGF处理25μg/m L 5-FU损伤的OP9细胞作为实验组,并设常规培养的OP9细胞作为空白对照组。分别用CCK-8法检测细胞存活率,流式细胞术(FCM)检测细胞凋亡,一氧化氮(NO)测试盒、一氧化氮合酶(NOS)测定试剂盒、Caspase-3活性检测试剂盒分别检测NO、i NOS、TNOS及Caspase-3的含量。结果与空白对照组比较,模型组细胞存活率明显降低(P<0.01);与模型组比较,EGF可以显著保护5-FU损伤的OP9细胞,使其存活率升高(P<0.01)。与空白对照组比较,模型组细胞早期凋亡率明显提高(P<0.05),Caspase-3含量明显增加(P<0.05),NO生成明显增多(P<0.05),i NOS表达明显上升(P<0.05);与模型组比较,实验组化疗损伤OP9细胞的早期凋亡率显著降低(P<0.05),Caspase-3含量明显下降(P<0.05),i NOS的表达明显受到抑制(P<0.01),NO的生成明显减少(P<0.05)。结论 EGF可能通过降低Caspase-3含量抑制细胞凋亡,减少NO的过度生成,发挥其细胞保护作用。展开更多
The research on stem cell cultures has attracted much attention due to the recent development of regenerative medicine. Therefore, higher functionalities for devices used for culturing cells are strongly demanded. In ...The research on stem cell cultures has attracted much attention due to the recent development of regenerative medicine. Therefore, higher functionalities for devices used for culturing cells are strongly demanded. In this study, we fabricated cell culture sheets using transparent polyimide (PI), parylene (PA), and polyetheretherketone (PEEK) to make polymer materials that had microstructures. We then cultured stromal marrow cells (OP9) on them and investigated the cell alignment within the microstructures. Hot embossing was used to fabricate the microstructures with a width and depth of 5 μm on the polymer substrates. Cultivation of the cells was confirmed on the transparent PI and PA sheets, however, it was not observed on the PEEK sheet. Slight alignment of the cells was also observed along with the microstructures.展开更多
基金supported by the National Natural Science Foundation (Nos 30871410 and 30600613)
文摘Mesenchymal stem cells （MSCs） are multipotent stem cells capable of differentiating into various cell types,including osteocytes,chondrocytes,adipocytes,myocytes,and tenocytes.However,the difficulty or failure in expanding the mouse MSCs in vitro greatly hampered important research in animal models.The OP9,a stromal cell line from mouse bone marrow,has hematopoietic supportive capacity.Here,we report that the OP9 has the immunophenotype （CD45-,CD11b-,FLK-1-,CD31-,CD34-,CD44＋,CD29＋,Sca-1＋,CD86-,and MHCII-） identical to canonical mouse MSCs.The expression of CD140a＋,CD140b＋,α-SMA＋ and Calponin＋ suggested the perivascular origin of OP9.Functionally,the OP9 had strong clonogenic ability and could be induced into osteocytes,chondrocytes and adipocytes.The lymphocyte transformation test （LTT） and mixed leukocyte reaction （MLR） showed that the OP9 could suppress T lymphocyte proliferation stimulated by nonspecific mitogens （PHA） or allogeneic lymphocytes （BALB/c T cells）.Finally,the migration of OP9 could be efficiently induced by bFGF,IGF-1,IL-3,PDGF-BB,TGF-β1 and TGF-β3.In conclusion,the OP9 were bona fide MSCs,and such homogenous cell line will be helpful to delineate biological features of MSCs at the stem cell level.
文摘The research on stem cell cultures has attracted much attention due to the recent development of regenerative medicine. Therefore, higher functionalities for devices used for culturing cells are strongly demanded. In this study, we fabricated cell culture sheets using transparent polyimide (PI), parylene (PA), and polyetheretherketone (PEEK) to make polymer materials that had microstructures. We then cultured stromal marrow cells (OP9) on them and investigated the cell alignment within the microstructures. Hot embossing was used to fabricate the microstructures with a width and depth of 5 μm on the polymer substrates. Cultivation of the cells was confirmed on the transparent PI and PA sheets, however, it was not observed on the PEEK sheet. Slight alignment of the cells was also observed along with the microstructures.
