Comparative gene expression analysis for pre-osteoblast MC3T3-E1 cells under non-adhesive culture toward osteocyte differentiation
Comparative gene expression analysis for pre-osteoblast MC3T3-E1 cells under non-adhesive culture toward osteocyte differentiation
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DOI：
10.1016/j.jbiosc.2021.09.004
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发表时间：
2021-11-13
影响因子：
2.8
通讯作者：
Adachi, Taiji
中科院分区：
文献类型：
--
作者：
Kim, Jeonghyun;Kigami, Hiroyuki;Adachi, Taiji
骨细胞在骨重建中起着重要的调节作用，又称终末分化细胞，起源于成骨细胞前体细胞，但其分化机制尚不清楚。鉴于目前尚未建立一种有效的体外诱导成骨细胞分化的方法，我们对小鼠成骨前细胞MC3T3-E1细胞进行了基因表达的比较分析，以阐明诱导成骨细胞分化的关键因素。在本研究中，我们通过调节细胞-底物相互作用和细胞-细胞相互作用来制备四种不同的培养环境：(I)贴壁培养模型上的低细胞密度和(Ii)高细胞密度；(Iii)非贴壁悬浮培养模型上的低细胞密度和高细胞密度。通过细胞-底物和细胞-细胞相互作用的比较，我们发现在非黏附漂浮培养条件下，细胞-底物相互作用的消除大大上调了成骨细胞前细胞中骨细胞标志物的基因表达。此外，在非粘附性球状细胞培养中，适度的细胞-细胞相互作用的存在进一步增强了成骨前细胞的骨细胞标志物的上调。这些结果提示了诱导前成骨细胞体外分化的最合适的培养环境。(C)2021年，日本生物技术学会。版权所有。
Osteocytes play an important role to modulate the bone remodeling and are also known as terminally differentiated cells originated from the osteoblast precursor cells, but its differentiation mechanism remains unclear. Since an efficient in vitro method to evoke the osteocyte differentiation from the osteoblast precursor cells has not been established, we conducted the comparative gene expression analysis for mouse pre-osteoblast MC3T3-E1 cells in order to elucidate the key factors to induce the osteocyte differentiation from the pre-osteoblast cells. In this study, we prepared four different culture environments by modulating their cell-substrate interaction and cell -cell interaction; (i) low and (ii) high cell density on the adhesive culture models, and (iii) low and (iv) high cell density on the non-adhesive floating culture models. By comparing these conditions in terms of cell-substrate and cell -cell interaction, we showed that the elimination of cell-substrate interaction under non-adhesive floating culture greatly up-regulated the gene expression of osteocyte markers in the pre-osteoblast cells. Moreover, the presence of moderate cell -cell interaction in the nonadhesive spheroid culture further enhanced the up-regulation of osteocyte markers for the pre-osteoblast cells. The results altogether suggest the most appropriate culture environment to induce the in vitro osteocyte differentiation of pre-osteoblast cells. (c) 2021, The Society for Biotechnology, Japan. All rights reserved.