YKL-40 Is Differentially Expressed in Human Embryonic Stem Cells and in Cell Progeny of the Three Germ Layers
YKL-40 Is Differentially Expressed in Human Embryonic Stem Cells and in Cell Progeny of the Three Germ Layers
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DOI：
10.1369/0022155411433331
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发表时间：
2012-03-01
影响因子：
3.2
通讯作者：
Mollgard, Kjeld
中科院分区：
文献类型：
--
作者：
Brochner, Christian B.;Johansen, Julia S.;Mollgard, Kjeld
分泌的糖蛋白 YKL-40 参与细胞分化、炎症和癌症进展。据报道，YKL-40 在人类早期发育过程中高表达，但其功能尚不清楚。将六种人胚胎干细胞 (hESC) 系在低或高氧张力的气氛中、在含有或不含碱性成纤维细胞生长因子的培养基中以及在包含小鼠胚胎成纤维细胞或人包皮成纤维细胞的饲养层上培养，以评估 hESC 及其后代是否产生 YKL-40 并表征分化过程中 YKL-40 的表达。通过酶联免疫吸附测定(ELISA)和定量RT-PCR测量分泌的YKL-40蛋白和YKL-40 mRNA表达。对连续切片集落进行 YKL-40 蛋白和多能 hESC（OCT4、NANOG）和胚层（HNF-3 beta、PDX1、CD34、p63、nestin、PAX6）标记物的染色。双标记显示 OCT4 阳性 hESC、PAX6 阳性神经外胚层细胞和 HNF-3 β 阳性内胚层细胞中 YKL-40 表达。分化的后代显示出强的YKL-40表达。在同一上皮样层内观察到 YKL-40 和 OCT4 阳性 hESC 与 YKL-40 阳性外胚层和神经外胚层谱系之间的突然转变。较深层的 YKL-40 阳性细胞缺乏与 OCT4 阳性细胞的接触。 YKL-40 在从 hESC 向外胚层和神经外胚层的初始细胞分化中可能很重要，并保留上皮形态，而后来分化为内胚层和中胚层则涉及向集落更深层的过渡。(J Histochem Cytochem 60: 188-204, 2012)
The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3 beta, PDX1, CD34, p63, nestin, PAX6) markers. Double-labeling showed YKL-40 expression in OCT4-positive hESCs, PAX6-positive neuroectodermal cells, and HNF-3 beta-positive endodermal cells. The differentiating progeny showed strong YKL-40 expression. Abrupt transition between YKL-40 and OCT4-positive hESCs and YKL-40-positive ecto- and neuroectodermal lineages was observed within the same epithelial-like layer. YKL-40-positive cells within deeper layers lacked contact with OCT4-positive cells. YKL-40 may be important in initial cell differentiation from hESCs toward ectoderm and neuroectoderm, with retained epithelial morphology, whereas later differentiation into endoderm and mesoderm involves a transition into the deeper layers of the colony.(J Histochem Cytochem 60: 188-204, 2012)