中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
1期
1-8
,共8页
干细胞%骨髓干细胞%富血小板血浆%骨髓间充质干细胞%软骨细胞%分化%Ⅱ型胶原%Ⅱ型胶原α1链%聚集蛋白聚糖%兔%关节软骨损伤%省级基金%干细胞图片文章
榦細胞%骨髓榦細胞%富血小闆血漿%骨髓間充質榦細胞%軟骨細胞%分化%Ⅱ型膠原%Ⅱ型膠原α1鏈%聚集蛋白聚糖%兔%關節軟骨損傷%省級基金%榦細胞圖片文章
간세포%골수간세포%부혈소판혈장%골수간충질간세포%연골세포%분화%Ⅱ형효원%Ⅱ형효원α1련%취집단백취당%토%관절연골손상%성급기금%간세포도편문장
背景:自体富血小板血浆激活后可释放多种生长因子,可以促进骨髓间充质干细胞的增殖与分化.目的:观察自体激活富血小板血浆对体外培养的兔骨髓间充质干细胞向成软骨细胞分化的影响.方法:取兔股骨骨髓,全骨髓贴壁法分离培养骨髓间充质干细胞;取第3代骨髓间充质干细胞,分别应用体积分数10%自体激活富血小板血浆和体积分数10%胎牛血清培养液进行体外培养,观察其向成软骨细胞分化情况.结果与结论:分离培养的兔骨髓间充质干细胞呈长梭形,传代后细胞生长迅速.流式细胞仪检测发现第3代细胞高表达CD29、CD44,而低表达CD45.免疫荧光细胞化学染色显示经自体激活富血小板血浆诱导的骨髓间充质干细胞表达Ⅱ型胶原;实时荧光定量PCR 检测发现经自体激活富血小板血浆诱导的骨髓间充质干细胞Ⅱ型胶原α1链基因和聚集蛋白聚糖基因表达明显高于经胎牛血清诱导的骨髓间充质干细胞(P <0.01).可见自体激活富血小板血浆具有促进兔骨髓间充质干细胞向软骨细胞方向分化的潜能.
揹景:自體富血小闆血漿激活後可釋放多種生長因子,可以促進骨髓間充質榦細胞的增殖與分化.目的:觀察自體激活富血小闆血漿對體外培養的兔骨髓間充質榦細胞嚮成軟骨細胞分化的影響.方法:取兔股骨骨髓,全骨髓貼壁法分離培養骨髓間充質榦細胞;取第3代骨髓間充質榦細胞,分彆應用體積分數10%自體激活富血小闆血漿和體積分數10%胎牛血清培養液進行體外培養,觀察其嚮成軟骨細胞分化情況.結果與結論:分離培養的兔骨髓間充質榦細胞呈長梭形,傳代後細胞生長迅速.流式細胞儀檢測髮現第3代細胞高錶達CD29、CD44,而低錶達CD45.免疫熒光細胞化學染色顯示經自體激活富血小闆血漿誘導的骨髓間充質榦細胞錶達Ⅱ型膠原;實時熒光定量PCR 檢測髮現經自體激活富血小闆血漿誘導的骨髓間充質榦細胞Ⅱ型膠原α1鏈基因和聚集蛋白聚糖基因錶達明顯高于經胎牛血清誘導的骨髓間充質榦細胞(P <0.01).可見自體激活富血小闆血漿具有促進兔骨髓間充質榦細胞嚮軟骨細胞方嚮分化的潛能.
배경:자체부혈소판혈장격활후가석방다충생장인자,가이촉진골수간충질간세포적증식여분화.목적:관찰자체격활부혈소판혈장대체외배양적토골수간충질간세포향성연골세포분화적영향.방법:취토고골골수,전골수첩벽법분리배양골수간충질간세포;취제3대골수간충질간세포,분별응용체적분수10%자체격활부혈소판혈장화체적분수10%태우혈청배양액진행체외배양,관찰기향성연골세포분화정황.결과여결론:분리배양적토골수간충질간세포정장사형,전대후세포생장신속.류식세포의검측발현제3대세포고표체CD29、CD44,이저표체CD45.면역형광세포화학염색현시경자체격활부혈소판혈장유도적골수간충질간세포표체Ⅱ형효원;실시형광정량PCR 검측발현경자체격활부혈소판혈장유도적골수간충질간세포Ⅱ형효원α1련기인화취집단백취당기인표체명현고우경태우혈청유도적골수간충질간세포(P <0.01).가견자체격활부혈소판혈장구유촉진토골수간충질간세포향연골세포방향분화적잠능.
@@@@ BACKGROUND:Platelet-rich plasma, when activated, could secret multiple growth factors which may promote the proliferation and differentiation of bone marrow-derived mesenchymal stem cel s. OBJECTIVE:To explore the effect of activated autologous platelet-rich plasma on the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cel s in vitro. METHODS:Rabbit bone marrow-derived mesenchymal stem cel s were isolated using the whole bone marrow adherence method. Passage 3 bone marrow-derived mesenchymal stem cel s were in vitro cultured with 10%activated autologous platelet-rich plasma and 10%fetal bovine serum separately. Chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cel s in vitro was observed. RESULTS AND CONCLUSION:Bone marrow-derived mesenchymal stem cel s were successful y isolated and exhibited a long-shuttle-shaped appearance. Cel s grew faster when passaged. Immunofluorescence staining showed that after induced by activated autologous platelet-rich plasma, bone marrow-derived mesenchymal stem cel s exhibited type Ⅱ col agen expression. Quantitative real-time PCR analysis showed that the expression ofα1 chain of type Ⅱ col agen and aggrecan mRNA was significantly up regulated in the bone marrow-derived mesenchymal stem cel s induced by activated autologous platelet-rich plasma than in the cel s induced by fetal bovine serum (P<0.01). Our findings in this study suggested that activated autologous platelet-rich plasma can promote the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cel s in vitro.
