中华细胞与干细胞杂志（电子版）
中華細胞與榦細胞雜誌（電子版）
중화세포여간세포잡지（전자판）
CHINESE JOURNAL OF CELL AND STEM CELL
2014年
1期
26-35
,共10页
全能干细胞%骨髓间充质干细胞%视网膜变性%细胞移植
全能榦細胞%骨髓間充質榦細胞%視網膜變性%細胞移植
전능간세포%골수간충질간세포%시망막변성%세포이식
Totipotent stem cell%Bone mesenchymal stem cells%Retinal degeneration%Cell transplantation
目的：检测多能成体祖细胞（MAPC）的培养条件对猴骨髓间充质细胞（BMMSCs）和人脂肪干细胞（hADSCs）生长的影响，旨在获得更适合治疗视网膜变性疾病的供体细胞。方法通过细胞形态观察、MTT实验、克隆形成率、PCR检测、以及成脂、成骨、成软骨分化潜能检测等，研究MAPC培养条件下猴BMMSCs和hADSCs的特征，并用DMEM/LG和MAPC培养条件培养的hADSCs进行RCS大鼠视网膜下腔移植，通过视网膜电图(ERG)和TUNEL检测，判断细胞移植治疗对视功能及视网膜细胞凋亡的影响。结果与常规培养基相比，MAPC培养条件能促进猴BMMSCs增殖，细胞变小，但传2代后，细胞变得宽大扁平，出现衰老征象；然而，MAPC培养条件下的hADSCs细胞增殖能力及克隆形成率均增强，形成的克隆较大可稳定传10代以上，且具有成脂、成骨、成软骨的多向分化潜能，细胞表面标记物及细胞因子出现差异表达：CD140b、CD90、CD47、HGF和PEDF显著上调,CD73、CD105和IL-6显著下调。与对照组相比，移植DMEM/LG和MAPC培养条件培养的hADSCs（P4）3周后，RCS大鼠的B波波幅明显升高，外核层细胞凋亡明显减少。结论 MAPC培养条件培养的hADSCs显示出更好的视网膜神经保护作用，适合用于治疗视网膜退行性疾病。
目的：檢測多能成體祖細胞（MAPC）的培養條件對猴骨髓間充質細胞（BMMSCs）和人脂肪榦細胞（hADSCs）生長的影響，旨在穫得更適閤治療視網膜變性疾病的供體細胞。方法通過細胞形態觀察、MTT實驗、剋隆形成率、PCR檢測、以及成脂、成骨、成軟骨分化潛能檢測等，研究MAPC培養條件下猴BMMSCs和hADSCs的特徵，併用DMEM/LG和MAPC培養條件培養的hADSCs進行RCS大鼠視網膜下腔移植，通過視網膜電圖(ERG)和TUNEL檢測，判斷細胞移植治療對視功能及視網膜細胞凋亡的影響。結果與常規培養基相比，MAPC培養條件能促進猴BMMSCs增殖，細胞變小，但傳2代後，細胞變得寬大扁平，齣現衰老徵象；然而，MAPC培養條件下的hADSCs細胞增殖能力及剋隆形成率均增彊，形成的剋隆較大可穩定傳10代以上，且具有成脂、成骨、成軟骨的多嚮分化潛能，細胞錶麵標記物及細胞因子齣現差異錶達：CD140b、CD90、CD47、HGF和PEDF顯著上調,CD73、CD105和IL-6顯著下調。與對照組相比，移植DMEM/LG和MAPC培養條件培養的hADSCs（P4）3週後，RCS大鼠的B波波幅明顯升高，外覈層細胞凋亡明顯減少。結論 MAPC培養條件培養的hADSCs顯示齣更好的視網膜神經保護作用，適閤用于治療視網膜退行性疾病。
목적：검측다능성체조세포（MAPC）적배양조건대후골수간충질세포（BMMSCs）화인지방간세포（hADSCs）생장적영향，지재획득경괄합치료시망막변성질병적공체세포。방법통과세포형태관찰、MTT실험、극륭형성솔、PCR검측、이급성지、성골、성연골분화잠능검측등，연구MAPC배양조건하후BMMSCs화hADSCs적특정，병용DMEM/LG화MAPC배양조건배양적hADSCs진행RCS대서시망막하강이식，통과시망막전도(ERG)화TUNEL검측，판단세포이식치료대시공능급시망막세포조망적영향。결과여상규배양기상비，MAPC배양조건능촉진후BMMSCs증식，세포변소，단전2대후，세포변득관대편평，출현쇠로정상；연이，MAPC배양조건하적hADSCs세포증식능력급극륭형성솔균증강，형성적극륭교대가은정전10대이상，차구유성지、성골、성연골적다향분화잠능，세포표면표기물급세포인자출현차이표체：CD140b、CD90、CD47、HGF화PEDF현저상조,CD73、CD105화IL-6현저하조。여대조조상비，이식DMEM/LG화MAPC배양조건배양적hADSCs（P4）3주후，RCS대서적B파파폭명현승고，외핵층세포조망명현감소。결론 MAPC배양조건배양적hADSCs현시출경호적시망막신경보호작용，괄합용우치료시망막퇴행성질병。
Objective To investigate the effects of multipotent adult progenitor cell (MAPC) conditions on biological characteristics and therapeutic effects of monkey bone marrow mesenchymal stem cells (BMMSCs) and human adipose-derived stem cells (hADSCs), thus to obtain optimal donor cells for treatment of retinal degeneration (RD). Methods The cell morphology, MTT assay, clone formation, adipogenic, osteogenic, chondrogenic differentiation potential and qRT-PCR technique were used to detect biological characteristics of monkey BMMSCs and hADSCs. The hADSCs were cultured under MAPC condition medium and transplanted into the subretinal space of RCS rats. The retinal functions were evaluated by electroretingram (ERG) recording, histological examination and TUNEL assay. Results Compared with conventional medium, MAPC conditions promoted the growth of monkey BMMSCs with smaller cell size. However, two passages later by subculture, monkey BMMSCs changed into larger and flatter morphology, showing senescence characteristics. MAPC culture conditions, compared with conventional hADSCs culture medium, maintained hADSC’s morphology, high proliferative capacity and multi-lineage differentiation potentials in long-term culture (beyond 10 passages), and fewer cells displayed senescence. Gene expression profiles for cell surface markers and cytokines showed a differential expression patterns:Under MAPC culture conditions, CD140b,CD90, CD47, HGF and PEDF were significantly up-regulated, whereas CD73, CD105 and IL-6 were significantly down-regulated compared with those under conventional culture medium. In comparison with untreated RCS rats, DMEM/LG-hADSCsand MAPC-hADSC transplanted RCS rats showed significantly improved b-wave amplitudes in ERG examination in 3 weeks after the transplantation, and significant reduction of apoptotic cells in the outer nuclear layer. Conclusions hADSCs treated with MAPC culture conditions were more effective in protect retinal neurons and hold great promise for clinical use in RD disease.
