杜丽丽1,2 林戈1,2 卢光琇1,2△.人诱导多能干细胞与人胚胎干细胞分化过程中Oct4/Nanog 基因
表达的比较[J].,2011,11(1):1-4 |
人诱导多能干细胞与人胚胎干细胞分化过程中Oct4/Nanog 基因
表达的比较 |
Comparison of Human Induced Pluripotent Stem Cells and Oct4/NanogGene Expression in Human Embryonic Stem Cell Differentiation |
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DOI: |
中文关键词: 诱导性多能干细胞 胚胎干细胞 分化 拟胚体 畸胎瘤 |
英文关键词: Induced pluripotent stem cells Embryonic stem cells Differentiation Embryoid bodies (EBs) Teratoma |
基金项目:国家973 项目(2005CB522705);国家863 计划项目(2006AA02A102) |
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中文摘要: |
目的:比较通过慢病毒方法获得的人诱导多能性干细胞(iPSCs)与人胚胎干细胞(hESCs)分化过程中全能性基因Oct4、
Nanog 的表达变化。方法:收集分化不同时间点的拟胚体(EBs),检测三胚层分化基因以及全能性基因Oct4/Nanog 的表达,并通
过畸胎瘤组织切片的荧光染色分析Oct4 的表达。结果:iPSCs 获得的EB 中内外三胚层分化基因表达的出现明显晚于hESCs 来
源的EB。不同于hESCs,iPSCs 悬浮培养获得的EBs 在体外培养18 天未见内源性Oct4、Nanog 基因表达的下调。未分化的iPSCs
注射严重联合免疫缺陷(SCID)小鼠培养10 周后获得的畸胎瘤中仍存在Oct4 阳性的细胞,但iPS-#2 中明显少于iPS-#5。结论:
通过慢病毒方法获得的iPSCs 虽然具有向三胚层分化的能力,但在分化过程中仍维持较高水平的全能性基因Oct4、Nanog 的表
达。 |
英文摘要: |
Objective: To compare the expression of pluripotency gene Oct4/Nanog in the differentiated human induced pluripotent
stem cells (iPSCs) and embryonic stem cells (ESCs). Methods: The RNA of embryoid bodies (EBs) derived from undifferentiated
human iPSCs and hESCs at different time of differentiation were collected. The gene expression related to three-layer differentiation and
pluripotency gene Oct4/Nanog was detected by RT-PCR. The Oct4 positive cells in the teratoma derived from both human iPSCs and
hESCs were detected by immunofluorescence staining. Results: The expression of endoderm and ectroderm related gene in the EBs
derived from iPSCs was later than that derived from hESCs. The expression of Oct4 and Nanog was still existence in D18 EBs derived
from undifferentiated human iPSCs. The Oct4 positive cells were detected in iPSCs derivded-teratoma. Conclusion: Although the iPSCs
can differentiate into three-layer cells, the higher expression of pluriotency gene Oct4/Nanog was maintained in the differentiated iPSCs
generated by lentivirus infection than hES. |
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