目的从基因水平了解中药在诱导大鼠骨髓间充质干细胞向神经细胞分化过程中的分子机制。方法应用基因芯片技术分析了黄芪(Radix Astragali Seu Hedysari,RASH)和天麻(Gastrodiaelata,GE)诱导大鼠骨髓间充质干细胞向神经元分化过程中差异...目的从基因水平了解中药在诱导大鼠骨髓间充质干细胞向神经细胞分化过程中的分子机制。方法应用基因芯片技术分析了黄芪(Radix Astragali Seu Hedysari,RASH)和天麻(Gastrodiaelata,GE)诱导大鼠骨髓间充质干细胞向神经元分化过程中差异表达的基因。结果在黄芪诱导大鼠骨髓间充质干细胞向神经元分化过程中,有96个差异表达基因(上调或下调倍数大于2);在天麻组有118个差异表达基因(上调或下调倍数大于2)。在两组中共发现与细胞分化相关的差异表达基因17个,其中差异表达明显的有3个,包括bHLH,Epiregulin,fibroblast growth factor 9。结论黄芪和天麻可有效地诱导骨髓间充质干细胞分化神经元,其作用机制涉及多种基因。展开更多
Background The two most basic properties of mesenchymal stem cells （MSCs） are the capacities to selfrenew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Whar...Background The two most basic properties of mesenchymal stem cells （MSCs） are the capacities to selfrenew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton＇ s Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton＇ s Jelly-derived MSCs to differentiate into nerve-like cells.Methods MSCs were cultured from the Wharton＇ s Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and [3-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy （EM）, and laser scanning confocal microscopy （LSCM） . The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction （RT-PCR）.Results MSCs in the Wharton＇ s Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease （GVHD）-related cells. Treatment with Salvia mihiorrhiza caused Wharton＇ s Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia mihiorrhiza and β-mercaptoethanol also induced MSCs to express nestin, β-tubulin Ⅲ, neurofilament （NF） and glial fibrillary acidic protein （GFAP）. It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed.Conclusions MSCs could be isolated from human umbilical cord Wharto展开更多
目的观察心脏营养素-1(CT-1)是否能促进经诱导分化剂5-氮杂胞苷(5-aza)诱导的骨髓间充质干细胞(BMMSCs)分化为心肌样细胞,并研究其相关机制。方法自成年大鼠骨髓中分离BMMSCs,分别以普通培养基(A组)、加入含CT-1的培养基(B组)培养、经5-...目的观察心脏营养素-1(CT-1)是否能促进经诱导分化剂5-氮杂胞苷(5-aza)诱导的骨髓间充质干细胞(BMMSCs)分化为心肌样细胞,并研究其相关机制。方法自成年大鼠骨髓中分离BMMSCs,分别以普通培养基(A组)、加入含CT-1的培养基(B组)培养、经5-aza诱导后加入普通培养基(C组)及5-aza加入含CT-1的培养基(D组)培养。观察细胞形态的改变,并通过免疫组化分析分化后细胞表达心脏特异性肌钙蛋白T(cTnT)的情况。电镜观察分化后细胞的超微结构及实时荧光定量检测α-actin、β-myosin heavy chain(β-MHC)、Nkx2.5、GATA4基因表达。结果C、D组的BMMSCs在培养4周后均形成心肌样细胞形态,并且均表达cTnT;D组BMMSCs分化的心肌样细胞形成了肌管样结构;D组α-actin、β-MHC、Nkx2.5、GATA4基因表达明显高于C组。结论CT-1可能通过对GATA4、Nkx2.5基因表达的调控而促进5-aza诱导的BMMSCs分化为心肌样细胞。展开更多
文摘Background The two most basic properties of mesenchymal stem cells （MSCs） are the capacities to selfrenew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton＇ s Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton＇ s Jelly-derived MSCs to differentiate into nerve-like cells.Methods MSCs were cultured from the Wharton＇ s Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and [3-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy （EM）, and laser scanning confocal microscopy （LSCM） . The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction （RT-PCR）.Results MSCs in the Wharton＇ s Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease （GVHD）-related cells. Treatment with Salvia mihiorrhiza caused Wharton＇ s Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia mihiorrhiza and β-mercaptoethanol also induced MSCs to express nestin, β-tubulin Ⅲ, neurofilament （NF） and glial fibrillary acidic protein （GFAP）. It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed.Conclusions MSCs could be isolated from human umbilical cord Wharto
文摘目的观察心脏营养素-1(CT-1)是否能促进经诱导分化剂5-氮杂胞苷(5-aza)诱导的骨髓间充质干细胞(BMMSCs)分化为心肌样细胞,并研究其相关机制。方法自成年大鼠骨髓中分离BMMSCs,分别以普通培养基(A组)、加入含CT-1的培养基(B组)培养、经5-aza诱导后加入普通培养基(C组)及5-aza加入含CT-1的培养基(D组)培养。观察细胞形态的改变,并通过免疫组化分析分化后细胞表达心脏特异性肌钙蛋白T(cTnT)的情况。电镜观察分化后细胞的超微结构及实时荧光定量检测α-actin、β-myosin heavy chain(β-MHC)、Nkx2.5、GATA4基因表达。结果C、D组的BMMSCs在培养4周后均形成心肌样细胞形态,并且均表达cTnT;D组BMMSCs分化的心肌样细胞形成了肌管样结构;D组α-actin、β-MHC、Nkx2.5、GATA4基因表达明显高于C组。结论CT-1可能通过对GATA4、Nkx2.5基因表达的调控而促进5-aza诱导的BMMSCs分化为心肌样细胞。
