Spontaneously Formed Spheroids from Mouse Compact Bone-Derived Cells Retain Highly Potent Stem Cells with Enhanced Differentiation Capability
Spontaneously Formed Spheroids from Mouse Compact Bone-Derived Cells Retain Highly Potent Stem Cells with Enhanced Differentiation Capability
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DOI：
10.1155/2019/8469012
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发表时间：
2019-01-01
影响因子：
4.3
通讯作者：
Kagami, Hideaki
中科院分区：
文献类型：
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作者：
Chen, Kai;Li, Xianqi;Kagami, Hideaki
我们最近的研究结果表明，球体的形成存在两种不同的机制，即自发机制和机械机制。本研究以自发形成的球体为研究对象，探讨了小鼠致密骨源性细胞(CBDC)自发形成的球体的特性。从(C57BL/6J)小鼠腿骨分离细胞，经酶消化后培养致密骨源性细胞。据报道，在特定水接触角的培养板上实现了自发球体的形成。用免疫荧光和定量逆转录聚合酶链式反应分析胚胎干细胞标志物的表达水平。然后，将来自球体的细胞诱导为成骨和神经样谱系。从CBDC自发形成的球体呈SSEA1、SOX2、Oct4和Nanog等ES细胞标志阳性。此外，岩藻糖基转移酶4/FUT4(SSEA1)、Sox2和Nanog的表达明显高于单层培养的细胞。间充质干细胞标志物在球体和单层培养细胞中的基因表达几乎相同，但SCA-1在球体中的表达较高。诱导后，类球形细胞成骨和神经性标志物的表达明显高于单层培养的细胞。自发形成的球体表达干细胞标记物，并显示出比传统单层培养的细胞更强的成骨和神经分化能力，这支持了优越的茎。
The results from our recent study showed the presence of two distinct spheroid-forming mechanisms, i.e., spontaneous and mechanical. In this study, we focused on the spontaneously formed spheroids, and the character of spontaneously formed spheroids from mouse compact bone-derived cells (CBDCs) was explored. Cells from (C57BL/6J) mouse leg bones were isolated, and compact bone-derived cells were cultured after enzymatic digestion. Spontaneous spheroid formation was achieved on a culture plate with specific water contact angle as reported. The expression levels of embryonic stem cell markers were analyzed using immunofluorescence and quantitative reverse transcription polymerase chain reaction. Then, the cells from spheroids were induced into osteogenic and neurogenic lineages. The spontaneously formed spheroids from CBDCs were positive for ES cell markers such as SSEA1, Sox2, Oct4, and Nanog. Additionally, the expressions of fucosyltransferase 4/FUT4 (SSEA1), Sox2, and Nanog were significantly higher than those in monolayer cultured cells. The gene expression of mesenchymal stem cell markers was almost identical in both spheroids and monolayer-cultured cells, but the expression of Sca-1 was higher in spheroids. Spheroid-derived cells showed significantly higher osteogenic and neurogenic marker expression than monolayer-cultured cells after induction. Spontaneously formed spheroids expressed stem cell markers and showed enhanced osteogenic and neurogenic differentiation capabilities than cells from the conventional monolayer culture, which supports the superior stemness.