脂肪干细胞生长特性研究Growth Characteristics of Human Adipose-Derived Stem Cells during Long Time Culture
姜丽丽,刘天庆,宋克东,关水,李香琴,葛丹
摘要(Abstract):
研究了脂肪干细胞在一个月连续培养中的生长增殖特性,及经长期连续培养后细胞的干细胞特性,同时探索了细胞周期蛋白表达与生长增殖特性之间的关系。分别采用CCK-8试剂盒与血细胞计数板来检测细胞活性与细胞数随时间的变化;用流式细胞仪检测细胞的死活情况;采用流式细胞仪和组织化学染色分析经一个月连续培养后细胞的干细胞特性;流式细胞仪和western blot分析不同时间段的细胞周期分布和细胞周期蛋白的表达。结果表明,脂肪干细胞在一个月的连续培养过程中始终保持很高的细胞活性和增殖能力。并且连续一个月培养后,仍然保持其干细胞特性。细胞周期蛋白A的增量表达,促进脂肪干细胞从S期向G2/M期过渡,使G2/M期细胞比率增加,处于分裂状态细胞含量增加,此时细胞处于快速增殖状态。因此,可以通过延长脂肪干细胞的传代培养时间来为细胞治疗提供更多高质量的移植用干细胞。并且在脂肪干细胞长期连续培养过程中,细胞周期蛋白A通过调节细胞周期分布而影响着细胞的生长增殖。
关键词(KeyWords): 脂肪干细胞;生长曲线;细胞周期;细胞周期蛋白A
基金项目(Foundation): 国家自然科学基金(31170945)
作者(Author): 姜丽丽,刘天庆,宋克东,关水,李香琴,葛丹
参考文献(References):
- [1]Le Blanc K,Rasmusson I,Sundberg B,et al.Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells[J].Lancet,2004,363(9419):1439-1441.
- [2]Schaechinger V,Erbs S,Elsaesser A,et al.Intracoronary bone marrow-derived progenitor cells in acute myocardial infarction[J].New England Journal of Medicine,2006,355(12):1210-1221.
- [3]Garcia-Olmo D,Garcia-Arranz M,Herreros D,et al.A phase I clinical trial of the treatment of Crohn's fistula by adipose mesenchymal stem cell transplantation[J].Diseases of the Colon&Rectum,2005,48(7):1416-1423.
- [4]Strem B M,Hicok K C,Zhu M,et al.Multipotential differentiation of adipose tissue-derived stem cells[J].The Keio Journal of Medicine,2005,54(3):132-141.
- [5]Safwani WKZW,Makpol S,Sathapan S,et al.The changes of stemness biomarkers expression in human adipose-derived stem cells during long-term manipulation[J].Biotechnology and Applied Biochemistry,2011,58(4):261-270.
- [6]Choi M R,Kim H Y,Park J Y,et al.Selection of optimal passage of bone marrow-derived mesenchymal stem cells for stem cell therapy in patients with amyotrophic lateral sclerosis[J].Neuroscience Letters,2010,472(2):94-98.
- [7]Kim J,Kang J,Park J,et al.Biological characterization of long-term cultured human mesenchymal stem cells[J].Archives of Pharmacal Research,2009,32(1):117-126.
- [8]Bonab M M,Alimoghaddam K,Talebian F,et al.Aging of mesenchymal stem cell in vitro[J].Bmc Cell Biology,2006,[2014-01-22].http://www.biomedcentral.com/1471-2121/7/14.
- [9]Zhu Y X,Liu T Q,Song K D,et al.Adipose-derived stem cell:a better stem cell than BMSC[J].Cell Biochemistry and Function,2008,26(6):664-675.
- [10]Cui L,Wu Y,Cen L,et al.Repair of articular cartilage defect in non-weight bearing areas using adipose derived stem cells loaded polyglycolic acid mesh[J].Biomaterials,2009,30(14):2683-2693.
- [11]Zuk P A,Zhu M,Mizuno H,et al.Multilineage cells from human adipose tissue:Implications for cell-based therapies[J].Tissue Eng,2001,7(2):211-228.
- [12]Xin Z F,Kim Y K,Jung S T.Risedronate inhibits human osteosarcoma cell invasion[J].J Exp Clin Cancer Res,2009,[2014-01-22].http://www.jeccr.com/content/28/1/105.
- [13]Ringden O,Uzunel M,Rasmusson I,et al.Mesenchymal stem cells for treatment of therapy-resistant graft-versus-host disease[J].Transplantation,2006,81(10):1390-1397.
- [14]Sekiya I,Larson B L,Smith J R,et al.Expansion of human adult stem cells from bone marrow stroma:Conditions that maximize the yields of early progenitors and evaluate their quality[J].Stem Cells,2002,20(6):530-541.
- [15]Yang X F,He X,He J,et al.High efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells[J].Journal of Biomedical Science,2011,[2014-01-22].http://www.jbiomedsci.com/content/18/1/59.
- [16]Ning H X,Liu G,Lin G T,et al.Identification of an aberrant cell line among human adipose tissue-derived stem cell isolates[J].Differentiation,2009,77(2):172-180.
- [17]Li X H,Du M,Liu X X,et al.Millimeter wave treatment promotes chondrocyte proliferation by upregulating the expression of cyclin-dependent kinase 2 and cyclin A[J].Int J Mol Med,2010,26(1):77-84.
- [18]Fei Q,Guo C,Xu X,et al.Osteogenic growth peptide enhances the proliferation of bone marrow mesenchymal stem cells from osteoprotegerin-deficient mice by CDK2/cyclin A[J].Acta Biochimica Et Biophysica Sinica,2010,42(11):801-806.