Human embryonic stem cells (hESC) can be induced to differentiate to trophoblast by bone morphogenetic proteins (BMPs) and by aggregation to form embryoid bodies (EB), but there are many differences and controversies ...Human embryonic stem cells (hESC) can be induced to differentiate to trophoblast by bone morphogenetic proteins (BMPs) and by aggregation to form embryoid bodies (EB), but there are many differences and controversies regarding the nature of the differentiated cells. Our goals herein were to determine if BG02 cells form trophoblast-like cells (a) in the presence of BMP4-plus-basic fibroblast growth factor (FGF-2) and (b) upon EB formation, and (c) whether the BMP4 antagonist noggin elicits direct effects on gene expression and hormone production in the cells. Transcriptome profiling of hESC incubated with BMP4/FGF-2 showed a down-regulation of pluripotency-associated genes, an up-regulation of trophoblast-associated genes, and either a down-regulation or no change in gene expression for many markers of the three embryonic germ layers. Yet, there was up-regulation of several genes associated with mesoderm, ectoderm, and endoderm, strongly suggesting that differentiation to trophoblast-like cells under the conditions used does not yield a homogeneous cell type. Several genes, heretofore unreported, were identified that are altered in hESC in response to BMP4-mediated differentiation. The production of human chorionic gonadotropin (hCG), progesterone, and estradiol in the differentiated cells confirmed that trophoblast-like cells were obtained. Gene expression by EB was characterized by an up-regulation of a number of genes associated with trophoblast, ectoderm, endoderm, and mesoderm, and the production of hCG and progesterone confirmed that trophoblast-like cells were formed. These results suggest that, in the presence of FGF-2, BG02 cells respond to BMP4 to yield trophoblast-like cells, which are also obtained upon EB formation. Thus, BMP4-mediated differentiation of hESC represents a viable cell system for studying early developmental events post-implantation;however, up-regulation of non-trophoblast genes suggests a somewhat diverse response to BMP4/FGF-2. Noggin altered the transcription of a limited number of genes but, not surprisingly, did not lead to secretion of hormones.展开更多
目前在体外研究中,主要通过分离胚体和骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)诱导这2种途径从人胚胎干细胞(hESC)分化获取滋养层细胞(TB)。胚体途径可基于细胞的黏附性和培养基中β-hCG的含量从胚体中分离获得TB,进而在...目前在体外研究中,主要通过分离胚体和骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)诱导这2种途径从人胚胎干细胞(hESC)分化获取滋养层细胞(TB)。胚体途径可基于细胞的黏附性和培养基中β-hCG的含量从胚体中分离获得TB,进而在三维培养体系中可检测细胞的侵袭性及细胞与基质的相互作用。BMP4途径通过去除外源性成纤维细胞生长因子及提高氧含量均可促进BMP4诱导TB的分化。虽然hESC分化TB模型目前还存在一些争议,但是相关研究可为探讨人类胚胎的植入及胎盘的形成提供重要的理论依据。展开更多
文摘Human embryonic stem cells (hESC) can be induced to differentiate to trophoblast by bone morphogenetic proteins (BMPs) and by aggregation to form embryoid bodies (EB), but there are many differences and controversies regarding the nature of the differentiated cells. Our goals herein were to determine if BG02 cells form trophoblast-like cells (a) in the presence of BMP4-plus-basic fibroblast growth factor (FGF-2) and (b) upon EB formation, and (c) whether the BMP4 antagonist noggin elicits direct effects on gene expression and hormone production in the cells. Transcriptome profiling of hESC incubated with BMP4/FGF-2 showed a down-regulation of pluripotency-associated genes, an up-regulation of trophoblast-associated genes, and either a down-regulation or no change in gene expression for many markers of the three embryonic germ layers. Yet, there was up-regulation of several genes associated with mesoderm, ectoderm, and endoderm, strongly suggesting that differentiation to trophoblast-like cells under the conditions used does not yield a homogeneous cell type. Several genes, heretofore unreported, were identified that are altered in hESC in response to BMP4-mediated differentiation. The production of human chorionic gonadotropin (hCG), progesterone, and estradiol in the differentiated cells confirmed that trophoblast-like cells were obtained. Gene expression by EB was characterized by an up-regulation of a number of genes associated with trophoblast, ectoderm, endoderm, and mesoderm, and the production of hCG and progesterone confirmed that trophoblast-like cells were formed. These results suggest that, in the presence of FGF-2, BG02 cells respond to BMP4 to yield trophoblast-like cells, which are also obtained upon EB formation. Thus, BMP4-mediated differentiation of hESC represents a viable cell system for studying early developmental events post-implantation;however, up-regulation of non-trophoblast genes suggests a somewhat diverse response to BMP4/FGF-2. Noggin altered the transcription of a limited number of genes but, not surprisingly, did not lead to secretion of hormones.
文摘目前在体外研究中,主要通过分离胚体和骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)诱导这2种途径从人胚胎干细胞(hESC)分化获取滋养层细胞(TB)。胚体途径可基于细胞的黏附性和培养基中β-hCG的含量从胚体中分离获得TB,进而在三维培养体系中可检测细胞的侵袭性及细胞与基质的相互作用。BMP4途径通过去除外源性成纤维细胞生长因子及提高氧含量均可促进BMP4诱导TB的分化。虽然hESC分化TB模型目前还存在一些争议,但是相关研究可为探讨人类胚胎的植入及胎盘的形成提供重要的理论依据。
