CAJ | 학술논문

目的从早期人胚分离和培养人早期胚胎间充质干细胞(hMSC),鉴定其生物学特性以及分化能力,观察hMSC在新生小鼠肾脏内存活和分布情况.方法取4～7周胚龄的人胚,分离组织进行培养,免疫组化和流式细胞术检测hMSC的生物学特性,以及成骨、成脂肪的分化能力.激光共聚焦显微镜观察PKH-26标记的hMSC在新生鼠肾脏的存活及分布情况.结果分离培养得到的hMSC呈纤维状,贴壁生长,增殖能力强.流式细胞仪检测显示细胞不表达CD34、CD45,但明显表达CD29、CD44、CD90.细胞免疫组化结果显示细胞表达SH-2、OCT-4.在成脂肪和成骨诱导培养的条件下具有多向分化的能力.观察1个月,PKH-26标记的hMSC在新生鼠肾脏中仍存活,10.0%±2.1%分布在肾小管组织.结论从早期人胚得到的hMSC有较强的增殖和分化能力,免疫力低,在小鼠肾脏内能艮期存活,并可能参与小管上皮细胞的发育.
목적 종조기인배분리화배양인조기배태간충질간세포(hMSC),감정기생물학특성이급분화능력,관찰hMSC재신생소서신장내존활화분포정황.방법 취4～7주배령적인배,분리조직진행배양,면역조화화류식세포술검측hMSC적생물학특성,이급성골、성지방적분화능력.격광공취초현미경관찰PKH-26표기적hMSC재신생서신장적존활급분포정황.결과 분리배양득도적hMSC정섬유상,첩벽생장,증식능력강.류식세포의검측현시세포불표체CD34、CD45,단명현표체CD29、CD44、CD90.세포면역조화결과현시세포표체SH-2、OCT-4.재성지방화성골유도배양적조건하구유다향분화적능력.관찰1개월,PKH-26표기적hMSC재신생서신장중잉존활,10.0%±2.1%분포재신소관조직.결론 종조기인배득도적hMSC유교강적증식화분화능력,면역력저,재소서신장내능간기존활,병가능삼여소관상피세포적발육.
Objective To determine the biological characteristics of human embryonic MSC (hMSCs) and their potential ability of differentiation, and to investigate the survival and distribution of hMSCs after transplantation into the kidney of newborn mice. Methods hMSCs were derived from 4-7 week-old embryos, then primary culture was done. The biological characteristics of hMSCs were detected by immunohistochemical methods and flow cytometry. Their differentiation potential was determined by coculture with conditioning medium. The survival and distribution of PKH-26-stained hMSCs in mice were observed by laser scanning confocal microscope. Results Flow cytometry and immunochemistry staining revealed that the expression of CD29, CD44, CD90, SH-2, OCT-4 was positive significantly, and CD34, CD45 was negative. The cells could be induced to differentiate to osteocytes and adipocytes under special conditions. After transplantation for 1 month, PKH-26-stained hMSCs still existed in the kidney of mice and co-localized in tubular epithelium by confocal microscope. Conclusion hMSCs derived from the early human embryo have the ability of proliferation and differentiation with low immunity, and may be involved in the development of renal tubule in newbem mice.