脂肪间充质干细胞相关分化与炎症相关.ppt
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1、脂肪间充质干细胞相关分化与炎症相关,威斯腾生物技术中心,威斯腾生物十二周年庆技术培训之,目录,脂肪间充质干细胞分离、培养和鉴定,脂肪间充质干细胞,骨关节炎和肥胖,脂肪间充质干细胞分化与炎症,脂肪间充质干细胞(ASCs).,Adipose tissue is recognized as a complex endocrine organ that plays central roles in energy homeostasis,feeding,insulin sensitivity,and inflammation.Adipose tissue contains different cell
2、types besides adipocytes,including adipose-derived stromal cells(ASCs).ASCs were first reported by Frohlich in 1972,as adherent,proliferating adipocyte precursors.Different names have been used to describe the adherent cell population that can be expanded from lipoaspirates,e.g.lipoblast,pericytes,p
3、readipocytes,processed lipoaspirates(PLA)cells,and many others.Several works demonstrated the stem cell-like plasticity of ASCs and their capability of differentiating into cells of mesodermal origin,such as adipocyte,osteocyte,chondrocyte and myocyte lineages.,Zuk PA,etc.Human adipose tissue is a s
4、ource of multipotent stem cells.Mol Biol Cell 2002;13:4279-4295,PLA,or,SVF(stromal vascular fraction),To clarify nomenclature,the International Fat Applied Technology Society(IFATS)proposed to name ASCs the isolated,plastic-adherent,multipotent cell population obtained from adipose tissue through di
5、fferent methods,Subcutaneous fat is an abundant and accessible source of both heterogeneous stromal vascular fraction(SVF)cells and a small number of relatively homogeneous ASCs.SVF cells include vascular endothelial cells and their progenitors,smooth muscle cells,immune regulatory monocytes/macroph
6、ages and T regulatory cells.,Caspar-Bauguil,S.,et al.,Adipose tissue lymphocytes:types and roles.J Physiol Biochem,2009.65(4):p.423-36.,Similarly to bone marrow mesenchymal stromal cells(BMSCs),ASCs were identifiedin vivo as cells with perivascular localization,in close contact with the other cell t
7、ypes.Like other MSC types,ASCs cannot be identified by a single specific marker,but multiple marker combinations are needed for the identification of the cell population.ASCs express the surface markers used for BMSC characterization,with some peculiarities.BMSCs express CD106 that is not or slightl
8、y expressed by ASCs;by contrast,BM-MSCs lack the expression of CD49d,which is expressed by ASCs.Following ex vivo expansion,some markers are normally expressed by ASCs,such as CD13,CD29,CD34,CD54,CD73,CD90,CD105 and MHC I.By contrast,markers of the angiogenic and haematopoietic lineages,such as CD14
9、,CD31,CD133,MHC II and CD45,are not or poorly expressed by ASCs.,Strem BM,Hicok KC,Zhu M,Wulur I,Alfonso Z,Schreiber RE,et al.Multipotential differentiation of adipose tissue-derived stem cells.Keio J Med 2005;54:13241,ASCs的分离培养,1)准备实验器材和试剂,置于超净工作台内紫外消毒等。2)取3周龄SD大鼠断颈处死,750ml/L乙醇浸泡5-10min,之后再更换酒精再浸泡5
10、分钟,最后移入超净工作台内。3)在超净工作台内严格遵循无菌操作要求。用组织剪剪开腹部皮肤直达到大腿内侧腹股沟处。更换组织剪和组织镊,分别取出腹股沟处的白色脂肪垫,置入含有双抗(含100U/mL青霉素和100g/mL链霉素)的PBS中。4)组织镊剔除白色脂肪块上的血管、淋巴结等其他组织。再用含双抗的PBS清洗3遍,置入青霉素瓶内剪碎为约1mm3大小的组织块。,5)将剪碎的组织块移入75mL超大培养瓶底部并涂布均匀,每瓶约放入0.4g脂肪组织块,将培养瓶翻转底部朝上,置入37、5%CO2 孵箱内贴壁1小时左右。6)肉眼观察培养瓶底部没有明显水滴的时候,贴壁小心加入510ml 的培养基(含改良型-M
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- 脂肪 间充质 干细胞 相关 分化 炎症
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