Effects of mouse fetal liver cell culture density on hematopoietic cell expansion in three-dimensional cocultures with stromal cells
Effects of mouse fetal liver cell culture density on hematopoietic cell expansion in three-dimensional cocultures with stromal cells
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DOI：
10.1177/0391398821996377
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发表时间：
2021-02
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影响因子：
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通讯作者：
Hirotoshi Miyoshi;Kenji Abo;Daiki Hosoya;K. Matsuo;Yoshio Utsumi
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文献类型：
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作者：
Hirotoshi Miyoshi;Kenji Abo;Daiki Hosoya;K. Matsuo;Yoshio Utsumi
目的：造血干细胞移植的广泛应用需要一种有效的体外扩增系统。在这项研究中，我们研究了培养密度对小鼠胎肝细胞（FLC）的影响，该细胞用作HC来源，用于在与两种小鼠基质细胞系（OP 9或C3 H10 T1/2）的三维（3D）共培养中扩增原始HC。材料与方法：将FLC以不同密度（1、2和10 × 107个细胞/cm 3）接种到有或无基质细胞层的多孔聚合物支架中，并在无外源性细胞因子的培养物中扩增HC 2周。结果：在共培养和单独FLC对照之间观察到培养密度对HC扩增的不同影响。在基质细胞共培养中，当FLC以低密度接种时，实现了HC的高度扩增。相反，对照中的扩增随着培养密度的增加而增强。对于FLC中存在的原始HC的扩增，与C3 H10 T1/2细胞的共培养上级与OP 9细胞的共培养，其中c-kit+造血祖细胞扩增29.3倍，CD 34+造血干细胞扩增8.3倍。在对照中，HC扩增低于任何共培养物，证明了共培养用于HC扩增的优点。结论：基质细胞系可用于在三维共培养中扩增来源于FLC的原始HC。培养密度是原始毛细胞有效扩增的关键因素，且这种效应因培养条件而异。
Objective: An effective ex vivo expansion system of primitive hematopoietic cells (HCs) is required for wider application of hematopoietic stem cell transplantation. In this study, we examined effects of culture density on mouse fetal liver cells (FLCs) used as an HC source for the expansion of primitive HCs in three-dimensional (3D) cocultures with two kinds of mouse stromal cell lines (OP9 or C3H10T1/2). Materials and methods: FLCs were seeded at different densities (1, 2, and 10 × 107 cells/cm3) into porous polymer scaffolds with or without stromal cell layers and HCs were expanded in the cultures for 2 weeks without exogenous cytokines. Results: Differential effects of culture density on HC expansion were observed between cocultures and solitary FLC controls. In stromal cell cocultures, high expansion of HCs was achieved when FLCs were seeded at low densities. In contrast, the expansion in the controls was enhanced with increasing culture densities. With respect to expansion of primitive HCs existing in the FLCs, cocultures with C3H10T1/2 cells were superior to those with OP9 cells with a 29.3-fold expansion for c-kit+ hematopoietic progenitor cells and 8.3-fold expansion for CD34+ hematopoietic stem cells. In the controls, HC expansion was lower than in any cocultures, demonstrating the advantages of coculturing for HC expansion. Conclusion: Stromal cell lines are useful in expanding primitive HCs derived from FLCs in 3D cocultures. Culture density is a pivotal factor for the effective expansion of primitive HCs and this effect differs by culture condition.