Expansion of mouse primitive hematopoietic cells in three-dimensional cultures on chemically fixed stromal cell layers
Expansion of mouse primitive hematopoietic cells in three-dimensional cultures on chemically fixed stromal cell layers
复制标题
化学固定基质细胞层三维培养中小鼠原始造血细胞的扩增
DOI：
10.1007/s10616-020-00417-4
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发表时间：
2020
期刊：
影响因子：
2.2
通讯作者：
Sugiyama Satoshi
中科院分区：
文献类型：
--
作者：
Miyoshi Hirotoshi;Shimizu Yuichiro;Yasui Yutaka;Sugiyama Satoshi
为了建立一个实用和方便的方法来扩大造血细胞（HC），我们应用化学固定的基质细胞层内形成的三维（3D）支架饲养的HC文化。使用两个连续培养物扩增HC。首先，将基质细胞在多孔聚合物支架内培养并形成组织工程构建体（TEC）;使用醛（甲醛或戊二醛）或有机溶剂（丙酮、甲醇或乙醇）固定含有基质细胞的支架。其次，将作为HC来源的小鼠胎肝细胞（FLC）在TEC上培养2周，并检查固定液对原始HC（c-kit+和CD 34+细胞）扩增的影响。在培养液中，用戊二醛固定的TEC上的原始HC扩增了2.5- 5.1倍，而用甲醛固定的TEC上的原始HC未扩增。然而，我们在使用有机溶剂固定的TEC的培养物中实现了原始HC> 5倍的扩增。在这些溶剂中，使用丙酮固定获得了最高的膨胀，大约为10倍。乙醇固定的TEC也支持原始HC的扩增（6.6- 8.0倍）。除了这些足够的扩展之外，固定TEC的程序和存储相当容易。因此，在化学固定的TEC上的HC扩增可能是用于扩增原始HC的实用方法。
To establish a practical and convenient method to expand hematopoietic cells (HCs), we applied chemically-fixed stromal cell layers formed within three-dimensional (3D) scaffolds to feeder of HC cultures. The HCs were expanded using two successive cultures. First, stromal cells were cultured within porous polymer scaffolds and formed tissue-engineered constructs (TECs); the scaffolds containing stromal cells, were fixed using aldehyde (formaldehyde or glutaraldehyde) or organic solvents (acetone, methanol or ethanol). Second, mouse fetal liver cells (FLCs), as a source of HCs, were cultured on the TECs for 2 weeks, and the effects of fixative solutions on expansion of primitive HCs (c-kit+and CD34+cells) were examined. In the cultures on aldehyde-fixed TECs, primitive HCs were expanded 2.5- to 5.1-fold in the cultures on TECs fixed with glutaraldehyde, whereas no expansions were detected in those fixed with formaldehyde. However, we achieved expansion of primitive HCs > fivefold in the cultures using TECs fixed with organic solvents. Among these solvents, the highest expansions—of roughly tenfold—were obtained using acetone fixation. Ethanol-fixed TECs also supported the expansion of the primitive HCs well (6.6- to 8.0-fold). In addition to these sufficient expansions, the procedure and storage of fixed TECs is fairly easy. Thus, HC expansion on chemically-fixed TECs may be a practical method for expanding primitive HCs.
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影响因子：
2.6
作者：
Chou, Song;Flygare, Johan;Lodish, Harvey F.
通讯作者：
Lodish, Harvey F.
多孔支架内固定的成纤维细胞的冷冻保存：预培养和支架胶原涂层对三维冷冻保存性能的影响
影响因子：
2.4
作者：
Miyoshi H;Ehashi T;Ohshima N;Jagawa A
通讯作者：
Jagawa A
影响因子：
2.6
作者：
Kawada, H;Ando, K;Hotta, T
通讯作者：
Hotta, T
影响因子：
3.3
作者：
Hirotoshi Miyoshi;M. Murao;N. Ohshima;T. Tun
通讯作者：
Hirotoshi Miyoshi;M. Murao;N. Ohshima;T. Tun
DOI：
10.1056/nejmoa1207285
发表时间：
2012-12-13
期刊：
The New England journal of medicine
影响因子：
--
作者：
de Lima M;McNiece I;Robinson SN;Munsell M;Eapen M;Horowitz M;Alousi A;Saliba R;McMannis JD;Kaur I;Kebriaei P;Parmar S;Popat U;Hosing C;Champlin R;Bollard C;Molldrem JJ;Jones RB;Nieto Y;Andersson BS;Shah N;Oran B;Cooper LJ;Worth L;Qazilbash MH;Korbling M;Rondon G;Ciurea S;Bosque D;Maewal I;Simmons PJ;Shpall EJ
通讯作者：
Shpall EJ