贫血小板血清对人羊膜间充质干细胞增殖分化的影响
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摘要
目的研究贫血小板血清(SPPP)对人羊膜间充质干细胞(hAMSCs)增殖分化的影响。方法利用酶消化法分离hAMSCs,制备SPPP培养hAMSCs,利用流式细胞术检测细胞表型,WST法绘制细胞生长曲线,诱导分化实验检测中胚层分化能力。结果 SPPP培养的hAMSCs表达CD73,CD90,CD105,不表达CD34,CD45,细胞倍增时间为72h,能够被诱导分化为骨,软骨,脂肪细胞。结论 SPPP作为hAMSCs的无动物源性成分培养基,为hAMSCs的细胞治疗及临床应用提供可能。
Objective To evaluate the effect of serum from platelet-poor plasma(SPPP) on the proliferation and differentiation of human amnion mesenchymal stem cells(hAMSCs). Methods hAMSCs were isolated by enzymatic digestion and cultured in SPPP, then immunophenotype of hAMSCs was examined by flow cytometry.The cell growth curves were measured by WST assay. Results hAMSCs cultured in SPPP expressed CD73,CD90 and CD105 positively, but not express CD34 and Cd45, doubling time of hAMSCs was approximately 72 h. The hAMSCs in SPPP medium were differentiated successfuly into osteoblasts, chondrocytes and adipocytes in appropriate induction medium. Conclusion SPPP has the potential to be used for the expansion of hAMSCs before clinical application.
Objective To evaluate the effect of serum from platelet-poor plasma(SPPP) on the proliferation and differentiation of human amnion mesenchymal stem cells(hAMSCs). Methods hAMSCs were isolated by enzymatic digestion and cultured in SPPP, then immunophenotype of hAMSCs was examined by flow cytometry.The cell growth curves were measured by WST assay. Results hAMSCs cultured in SPPP expressed CD73,CD90 and CD105 positively, but not express CD34 and Cd45, doubling time of hAMSCs was approximately 72 h. The hAMSCs in SPPP medium were differentiated successfuly into osteoblasts, chondrocytes and adipocytes in appropriate induction medium. Conclusion SPPP has the potential to be used for the expansion of hAMSCs before clinical application.
引文
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[2]Koellensperger E,Bollinger N,Dexheimer V,et al.Choosing the right type of serum for different applications of human adipose tissue-derived stem cells:influence on proliferation and differentiation abilities[J].Cytotherapy,2014,16(6):789-799.
[3]Yang L,Ma J,Gan S,et al.Platelet poor plasma gel combined with amnion improves the therapeutic effects of human umbilical cord-derived mesenchymal stem cells on wound healing in rats[J].Mol Med Rep,2017,16(3):3494-3502.
[4]Chisini LA,Karam SA,Noronha TG,et al.Platelet-Poor Plasma as a Supplement for Fibroblasts Cultured in Platelet-Rich Fibrin[J].Acta Stomatol Croat,2017,51(2):133-140.
[5]Shahidi M,Vatanmakanian M,Arami MK,et al.A comparative study between platelet-rich plasma and platelet-poor plasma effects on angiogenesis[J].Med Mol Morphol,2018,51(1):21-31.
[6]Batista JD,Justino Oliveira Limirio PH,Rocha FS,et al.Influence of platelet-poor plasma on angiogenesis and maintenance of volume in autogenous bone grafts[J].J Oral Maxillofac Surg,2018,2391(18):30594-30599.
[7]Franco D,Franco T,Schettino AM,et al.Protocol for obtaining platelet-rich plasma(PRP),platelet-poor plasma(PPP),and thrombin for autologous use[J].Aesthetic Plast Surg,2012,36(5):1254-1259.
[8]Sen F,Pinarba?i B,I?sever H,et al.Postprandial platelet-poor plasma 5-hydroxytryptamine concentrations during diarrhea and constipation periods of alternatingtype irritable bowel syndrome patients[J].Turk J Gastroenterol,2011,22(3):270-278.
[9]Woo SH,Kim JP,Park JJ,et al.Autologous platelet-poor plasma gel for injection laryngoplasty[J].Yonsei Med J,2013,54(6):1516-1523.