人胚胎干细胞来源的间充质干细胞（hES-MSCs）的最佳低氧预处理及其特性
Optimal Hypoxic Preconditioning of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells (hES-MSCs) and Their Characteristics.
作者信息
Lee Seung Min, Jun Dae Won, Kang Hyeon Tae, Oh Ju Hee, Saeed Waqar Khalid, Ahn Sang Bong
机构信息
Department of Translational Medicine, Graduate School of Biomedical Science & Engineering, Hanyang University, Seoul, Korea.
Department of Internal Medicine, Hanyang University School of Medicine, Seoul, Korea.
出版信息
Int J Stem Cells. 2021 May 30;14(2):221-228. doi: 10.15283/ijsc20096.
BACKGROUND AND OBJECTIVES
Hypoxia is frequently used to enhance stem cell function. However, the optimal level of hypoxia for growth and function of human embryonic stem cell-derived mesenchymal stem cells (hES-MSCs) is yet to be determined. The purpose of this study was to find the optimal level of hypoxia for hES-MSCs and characteristics of hES-MSCs cultured under these optimal hypoxic conditions.
METHODS AND RESULTS
Cell viability and changes in the morphology of hES-MSCs were determined through cell proliferation and CCK-8 assay. The hES-MSCs were preconditioned under various hypoxic conditions (0.5∼5% O and 24∼72 h). The expression of cytokines in each culture condition was compared using cytokine array analysis. The morphology of hES-MSCs did not change under various hypoxic culture conditions. hES-MSCs viability after 48 h incubation in 2% O condition was higher than that in normoxic condition. HIF1 expression was increased up to six folds after 48 h of hypoxic preconditioning. HIF1 expression in hES-MSCs peaked after 48 h of incubation in 1% O condition. The expressions of PDGF-BB, IGFBP-6, VEGF-A, and angiogenin were increased after hES-MSCs were incubated for 48 h in 2% O condition.
CONCLUSIONS
The hES-MSCs viability and expressions of PDGF-BB, IGFBP-6, VEGF-A, and angiogenin increased after 48 h incubation in 2% O condition.
背景与目的
缺氧常被用于增强干细胞功能。然而，人胚胎干细胞来源的间充质干细胞（hES-MSCs）生长和功能的最佳缺氧水平尚未确定。本研究的目的是找出hES-MSCs的最佳缺氧水平以及在这些最佳缺氧条件下培养的hES-MSCs的特性。
方法与结果
通过细胞增殖和CCK-8检测来确定hES-MSCs的细胞活力和形态变化。将hES-MSCs在各种缺氧条件（0.5∼5% O₂和24∼72小时）下进行预处理。使用细胞因子阵列分析比较每种培养条件下细胞因子的表达。在各种缺氧培养条件下，hES-MSCs的形态没有变化。在2% O₂条件下孵育48小时后，hES-MSCs的活力高于常氧条件下的活力。缺氧预处理48小时后，HIF1表达增加至六倍。在1% O₂条件下孵育48小时后，hES-MSCs中HIF1表达达到峰值。在2% O₂条件下将hES-MSCs孵育48小时后，PDGF-BB、IGFBP-6、VEGF-A和血管生成素的表达增加。
结论
在2% O₂条件下孵育48小时后，hES-MSCs的活力以及PDGF-BB、IGFBP-6、VEGF-A和血管生成素的表达增加。
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