CAJ | 학술논문

背景:血小板裂解液是浓缩后血小板的裂解产物,含有多种活性成分.有研究表明这些活性成分可促进间充质干细胞的增殖和分化,但血小板裂解液作为一个整体,对间充质干细胞成骨分化的作用尚不明确.目的:分析血小板裂解液对人脐带间充质干细胞成骨诱导分化的干预效应.方法:采用胶原酶消化法分离人脐带间充质干细胞,体外培养扩增,流式细胞仪进行细胞表型鉴定.实验分为以下4组:普通矿化液组,血小板裂解液组,矿化液-血小板裂解液联合诱导组,对照组.加入相应诱导培养基诱导培养2周.结果与结论:分离得到的人脐带间充质干细胞的细胞表型CD34(-)﹑CD45(-)、HLA-DR(-),CD44(+)﹑CD105(+)﹑CD146(+).茜素红染色在3个诱导组中皆为阳性,染色效果未见明显差异,而对照组阴性.碱性磷酸酶活性测定显示3个诱导组的碱性磷酸酶活性都明显高于对照组(P<0.05),矿化液-血小板裂解液联合诱导组碱性磷酸酶活性明显高于普通矿化液组和血小板裂解液组(P<0.05).结果显示在体外条件下,单纯的5%血小板裂解液可以诱导人脐带间充质干细胞向成骨细胞定向分化,且联合应用矿化液时能更有效地诱导人脐带间充质干细胞分化为成骨细胞.
배경:혈소판렬해액시농축후혈소판적렬해산물,함유다충활성성분.유연구표명저사활성성분가촉진간충질간세포적증식화분화,단혈소판렬해액작위일개정체,대간충질간세포성골분화적작용상불명학.목적:분석혈소판렬해액대인제대간충질간세포성골유도분화적간예효응.방법:채용효원매소화법분리인제대간충질간세포,체외배양확증,류식세포의진행세포표형감정.실험분위이하4조:보통광화액조,혈소판렬해액조,광화액-혈소판렬해액연합유도조,대조조.가입상응유도배양기유도배양2주.결과여결론:분리득도적인제대간충질간세포적세포표형CD34(-)﹑CD45(-)、HLA-DR(-),CD44(+)﹑CD105(+)﹑CD146(+).천소홍염색재3개유도조중개위양성,염색효과미견명현차이,이대조조음성.감성린산매활성측정현시3개유도조적감성린산매활성도명현고우대조조(P<0.05),광화액-혈소판렬해액연합유도조감성린산매활성명현고우보통광화액조화혈소판렬해액조(P<0.05).결과현시재체외조건하,단순적5%혈소판렬해액가이유도인제대간충질간세포향성골세포정향분화,차연합응용광화액시능경유효지유도인제대간충질간세포분화위성골세포.
BACKGROUND:Platelet lysate is the pyrolysis products of concentrated platelet which contains many active ingredients. Studies have shown that the active ingredients can promote the proliferation and differentiation of mesenchymal stem cel s, but the effect of platelet lysate on the osteogenic differentiation of mesenchymal stem cel s is not clear. OBJECTIVE:To study the intervention effect of platelet lysate in osteogenic differentiation of human umbilical cord derived mesenchymal stem cel s in vitro. METHODS:Human umbilical cord derived mesenchymal stem cel s isolated by col agenase digesting were cultured and amplified in vitro. Flow cytometer was used to detect the cel surface markers. The experiment was divided into four groups:mineralization medium group, platelet lysate group, mineralization medium-platelet lysate group and control group. The cel s were cultured with corresponding induction medium for 2 weeks, then alizarin red staining was used to identify the oxteoblast, and alkaline phosphatase activity was used to compare the differentiation activity. RESULTS AND CONCLUSION:Flow cytometer results showed that CD34, CD45 and HLA-DR were negative in the human umbilical cord derived mesenchymal stem cel s while CD44, CD105 and CD146 were positive. Alizarin red staining was positive in three induced groups with no obvious differences, and it was negative in the control group. The alkaline phosphatase activity level in three induced groups was significantly higher than that in the control group (P<0.05), and the alkaline phosphatase activity level in mineralization medium-platelet lysate group was significantly higher than that in the mineralization medium group and platelet lysate group (P<0.05). Results show that 5%platelet lysate alone can induce human umbilical cord derived mesenchymal stem cel s to differentiate into osteoblasts in vitro. The induction wil be more efficient when combining 5%platelet lysate with mineralization medium.