Controlling fluid flow to improve cell seeding uniformity.
Controlling fluid flow to improve cell seeding uniformity.
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DOI：
10.1371/journal.pone.0207211
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Gadegaard N
中科院分区：
文献类型：
--
作者：
Reynolds PM;Holzmann Rasmussen C;Hansson M;Dufva M;Riehle MO;Gadegaard N
Standard methods for seeding monolayer cell cultures in a multiwell plate or dish do not uniformly distribute cells on the surface. With traditional methods, users find aggregation around the circumference, in the centre, or a combination of the two. This variation is introduced due to the macro scale flow of the cell seeding suspension, and movement of the dish before cells can settle and attach to the surface. Reproducibility between labs, users, and experiments is hampered by this variability in cell seeding. We present a simple method for uniform and user-independent the cell seeding using an easily produced uniform cell seeder (UCS) device. This allows precise control of cell density in a reproducible manner. By containing the cell seeding suspension in a defined volume above the culture surface with the UCS, fluctuations in cell density are minimised. Seeding accuracy, as defined by the actual cell density versus the target seeding density is improved dramatically across users with various levels of expertise. We go on to demonstrate the impact of local variation in cell density on the lineage commitment of human embryonic stem cells (hESCs) towards pancreatic endoderm (PE). Variations in the differentiation profile of cells across a culture well closely mirror variations in cell density introduced by seeding method–with the UCS correcting variations in differentiation efficiency. The UCS device provides a simple and reproducible method for uniform seeding across multiple culture systems.
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影响因子：
3.7
作者：
Kim DS;Lee MW;Yoo KH;Lee TH;Kim HJ;Jang IK;Chun YH;Kim HJ;Park SJ;Lee SH;Son MH;Jung HL;Sung KW;Koo HH
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Koo HH
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Lee CM;Hu J
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14
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Lee LC;Gadegaard N;de Andrés MC;Turner LA;Burgess KV;Yarwood SJ;Wells J;Salmeron-Sanchez M;Meek D;Oreffo RO;Dalby MJ
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Dalby MJ
影响因子：
17.3
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Fernandes TG;Diogo MM;Clark DS;Dordick JS;Cabral JM
通讯作者：
Cabral JM
影响因子：
1.2
作者：
Whyte, Jemima L.;Ball, Stephen G.;Shuttleworth, C. Adrian;Brennan, Keith;Kielty, Cay M.
通讯作者：
Kielty, Cay M.