CAJ | 학술논문

目的对人脐带Wharton胶间质干细胞(human umbilical cord Wharton's Jelly-derived MSC,hWJMSC)进行免疫特性的体外和体内研究,探讨其应用于临床的可行性.方法取人脐带并分离Wharton胶,组织块法培养hWJMSC,通过细胞表面干细胞标志和多向分化实验对其进行鉴定;通过流式细胞术、免疫组织化学及RT-PCR等实验方法体外检测hWJMSC的主要组织相容性抗原(HLA-ABC和HLA-DPDQDR,即MHC-Ⅰ和MHC-Ⅱ)、表面共刺激分子(CD40、CD80、CD86)和免疫抑制分子(HLAC、IDO、PGE2)等的表达;通过细胞因子蛋白芯片和Western blot实验检测并验证hWJMSC中与免疫抑制相关的细胞因子(IL-10、TGF-β、VEGF、HGF)的表达;另外,还构建了hWJMSC-支架复合体,将其埋人家兔背部皮下,观察其在体内的免疫反应.结果从人脐带Wharton胶中分离、培养获得长梭形细胞,经流式细胞术及多向分化实验鉴定其为间质干细胞;流式细胞术及免疫组化等检测显示所分离的hWJMSC不表达MHC-Ⅱ,表达MHC-Ⅰ,阳性表达免疫抑制因子HLA-G、IDO、PGE2以及免疫抑制相关细胞因子IL-10、TGF-β、VEGF、HGF,表明其具有低免疫原性和诱导免疫耐受的能力;hWJMSC-支架埋人家兔皮下未引起免疫反应.结论 hWJMSC的免疫原性低,具有抑制免疫排斥、诱导宿主免疫耐受的能力,可应用于异体移植.
목적 대인제대Wharton효간질간세포(human umbilical cord Wharton's Jelly-derived MSC,hWJMSC)진행면역특성적체외화체내연구,탐토기응용우림상적가행성.방법 취인제대병분리Wharton효,조직괴법배양hWJMSC,통과세포표면간세포표지화다향분화실험대기진행감정;통과류식세포술、면역조직화학급RT-PCR등실험방법체외검측hWJMSC적주요조직상용성항원(HLA-ABC화HLA-DPDQDR,즉MHC-Ⅰ화MHC-Ⅱ)、표면공자격분자(CD40、CD80、CD86)화면역억제분자(HLAC、IDO、PGE2)등적표체;통과세포인자단백심편화Western blot실험검측병험증hWJMSC중여면역억제상관적세포인자(IL-10、TGF-β、VEGF、HGF)적표체;령외,환구건료hWJMSC-지가복합체,장기매인가토배부피하,관찰기재체내적면역반응.결과 종인제대Wharton효중분리、배양획득장사형세포,경류식세포술급다향분화실험감정기위간질간세포;류식세포술급면역조화등검측현시소분리적hWJMSC불표체MHC-Ⅱ,표체MHC-Ⅰ,양성표체면역억제인자HLA-G、IDO、PGE2이급면역억제상관세포인자IL-10、TGF-β、VEGF、HGF,표명기구유저면역원성화유도면역내수적능력;hWJMSC-지가매인가토피하미인기면역반응.결론 hWJMSC적면역원성저,구유억제면역배척、유도숙주면역내수적능력,가응용우이체이식.
Objective To probe the immunological traits of mesenchymal stem cells derived from umbilical cord Wharton's jelly (WJMSCs). Methods The diced Wharton's jelly which was from healthy fullterm birth human umbilical cord was cultured. The mesenchymal stem cells were identified with mesenchymal stem cells markers expression by flow cytometry and multiple differentiation ability. The expression of MHC- Ⅰ / Ⅱ, costimulatory molecules (CD40, CD80 and CD86) was detected with flow cytomctry, immunocytochemistry, and RT-PCR. The expression of immune inhibitors like HLA-G, IDO, and PGE2 was detected by immunocytochemistry and RT-PCR. The expression of immune-related molecules as IL-10, TGF-β, FGF and VEGF was detected with antibody microarray and western blot. Further more, to clarify the in vivo immune reaction of hWJMSCs, we fabricated the hWJMSC-scaffold constructs and implanted them into the rabbit backs. The lymphocyte infiltration and implanted cell survival observed with immunofluorescence. Results After culturinge of diced Wharton's jelly tissue, we obtained spindle-shaped cells. With differentiation medium, the cells can differentiate into osteoblasts, chongdrocytes, adipose cells and schwann cells. Expression of MHC, costimulatory molecules, and a series of immune suppressive-related molecules was found. Immune inhibitors as HLA-G, 1DO, PGE2, and immune suppressive related molecules as HGF, VEGF, TGFand IL-10 were positively expressed. But the cells did not express MHC-Ⅱ. No immune rejection was observed in vivo after implantation of hWJMSC-scaffold constructs. Conclusion It can be concluded that hWJMSCs have very low immunogenicity, which means the cells have potential to induce immune tolerance.The hWJMSCs do not provoke immune rejection in vivo.