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
45期
7834-7839
,共6页
韦葛堇%黄育强%覃万安%廖长川%林舟丹
韋葛堇%黃育彊%覃萬安%廖長川%林舟丹
위갈근%황육강%담만안%료장천%림주단
干细胞%骨髓干细胞%骨髓间充质干细胞%细胞共培养%诱导分化%转化生长因子β1%Ⅱ型胶原%髓核细胞%类髓核细胞%省级基金%干细胞图片文章
榦細胞%骨髓榦細胞%骨髓間充質榦細胞%細胞共培養%誘導分化%轉化生長因子β1%Ⅱ型膠原%髓覈細胞%類髓覈細胞%省級基金%榦細胞圖片文章
간세포%골수간세포%골수간충질간세포%세포공배양%유도분화%전화생장인자β1%Ⅱ형효원%수핵세포%류수핵세포%성급기금%간세포도편문장
背景:骨髓间充质干细胞近年来作为种子细胞被广泛应用于髓核组织工程。在诱导骨髓间充质干细胞向髓核细胞分化的过程中,需要提供适当的细胞外微环境以及生长因子。
<br> 目的:研究体外与髓核细胞非接触共培养和生物诱导剂条件下,兔骨髓间充质干细胞向类髓核样软骨细胞分化的可行性。
<br> 方法:用单层培养法分别分离培养兔骨髓间充质干细胞和髓核细胞。选用第3代的髓核细胞和骨髓间充质干细胞置于Transwel 培养皿上下层构建共培养体系并加入转化生长因子β1为主的生物诱导剂诱导培养21 d,以高糖DMEM培养基单独培养的骨髓间充质干细胞作为阴性对照。
<br> 结果与结论:分离培养的兔骨髓间充质干细胞细胞表面CD29和CD44表达阳性,CD34、CD45的细胞表达阴性。与髓核细胞共培养诱导21 d后,可观察到骨髓间充质干细胞形态向多角形类圆形转变,胞质中分泌Ⅱ型胶原颗粒明显增多,RT-PCR结果表明诱导后细胞中Ⅱ型胶原mRNA表达明显增高。结果说明骨髓间充质干细胞在与髓核细胞共培养加生物诱导剂条件下可成功诱导分化为类髓核细胞。
揹景:骨髓間充質榦細胞近年來作為種子細胞被廣汎應用于髓覈組織工程。在誘導骨髓間充質榦細胞嚮髓覈細胞分化的過程中,需要提供適噹的細胞外微環境以及生長因子。
<br> 目的:研究體外與髓覈細胞非接觸共培養和生物誘導劑條件下,兔骨髓間充質榦細胞嚮類髓覈樣軟骨細胞分化的可行性。
<br> 方法:用單層培養法分彆分離培養兔骨髓間充質榦細胞和髓覈細胞。選用第3代的髓覈細胞和骨髓間充質榦細胞置于Transwel 培養皿上下層構建共培養體繫併加入轉化生長因子β1為主的生物誘導劑誘導培養21 d,以高糖DMEM培養基單獨培養的骨髓間充質榦細胞作為陰性對照。
<br> 結果與結論:分離培養的兔骨髓間充質榦細胞細胞錶麵CD29和CD44錶達暘性,CD34、CD45的細胞錶達陰性。與髓覈細胞共培養誘導21 d後,可觀察到骨髓間充質榦細胞形態嚮多角形類圓形轉變,胞質中分泌Ⅱ型膠原顆粒明顯增多,RT-PCR結果錶明誘導後細胞中Ⅱ型膠原mRNA錶達明顯增高。結果說明骨髓間充質榦細胞在與髓覈細胞共培養加生物誘導劑條件下可成功誘導分化為類髓覈細胞。
배경:골수간충질간세포근년래작위충자세포피엄범응용우수핵조직공정。재유도골수간충질간세포향수핵세포분화적과정중,수요제공괄당적세포외미배경이급생장인자。
<br> 목적:연구체외여수핵세포비접촉공배양화생물유도제조건하,토골수간충질간세포향류수핵양연골세포분화적가행성。
<br> 방법:용단층배양법분별분리배양토골수간충질간세포화수핵세포。선용제3대적수핵세포화골수간충질간세포치우Transwel 배양명상하층구건공배양체계병가입전화생장인자β1위주적생물유도제유도배양21 d,이고당DMEM배양기단독배양적골수간충질간세포작위음성대조。
<br> 결과여결론:분리배양적토골수간충질간세포세포표면CD29화CD44표체양성,CD34、CD45적세포표체음성。여수핵세포공배양유도21 d후,가관찰도골수간충질간세포형태향다각형류원형전변,포질중분비Ⅱ형효원과립명현증다,RT-PCR결과표명유도후세포중Ⅱ형효원mRNA표체명현증고。결과설명골수간충질간세포재여수핵세포공배양가생물유도제조건하가성공유도분화위류수핵세포。
BACKGROUND:Bone marrow mesenchymal stem cells in recent years have been widely used as seed cells for nucleus pulposus tissue engineering. Appropriate extracellular microenvironment and growth factors are required for the induction of bone marrow mesenchymal stem cells differentiating into nucleus pulposus cells.
<br> OBJECTIVE:To observe the differentiation of rabbit bone marrow mesenchymal stem cells into nucleus pulposus-like cells in vitro induced by biological inducers and co-culture conditions.
<br> METHODS:Rabbit bone marrow mesenchymal stem cells and nucleus pulposus cells were isolated and cultured using monolayer culture method, respectively. Passage 3 bone marrow mesenchymal stem cells were placed in the Transwel culture plates for co-culture with the passage 3 nucleus pulposus cells and then were induced biological y to differentiate into the nucleus pulposus-like cells for 21 days. Bone marrow mesenchymal stem cells were cultured alone in the high-glucose Dulbecco’s modified Eagle’s medium as negative controls.
<br> RESULTS AND CONCLUSION:The expressions of CD29 and CD44 were positive and CD34 and CD45 were negative in the bone marrow mesenchymal stem cells. After 21 days of co-culture with nucleus pulposus cells, the morphology of bone marrow mesenchymal stem cells changed obviously from polygonal to oval shape, and col agen type Ⅱ granules were increased significantly. Reverse transcription-PCR results showed that expression of col agen type Ⅱ mRNA was significantly increased after induction. These findings indicate that bone marrow mesenchymal stem cells can be induced into differentiate into nucleus pulposus-like cells in co-culture plus biological inducer conditions in vitro.