Szaraz Peter, Librach Matthew, Mander Poonam, Hoseini Banafshe, Librach Max, Iqbal Farwah, Librach Clifford
Create Program Inc., Suite 412, Toronto, Ontario M5G 1N8, Canada
Create Program Inc., Suite 412, Toronto, Ontario M5G 1N8, Canada.
Biol Open. 2019 Jul 25;8(7):bio045294. doi: 10.1242/bio.045294.
High quality cell cultures require reliable laboratory practices. Today's small-scale cell culture format is dominated by circular topology vessels, with the inherent disadvantage of secondary flow induced each time the cell cultures are repositioned. The secondary flow generates uneven sedimentation and adherence that negatively impacts cell culture quality. Here we show a modification of the circular culture vessel that abrogates these disturbances. Cell culture wells were augmented with a central column to diminish secondary flow. Human carcinoma cell lines (BeWo, JEG-3), mesenchymal stem cells [human umbilical cord perivascular cells (HUCPVC)] and mouse embryonic fibroblasts (MEF) were cultured in both column-augmented and regular culture wells. Human carcinoma cell cultures showed even cell densities and significantly more viable cells in column-augmented vessels. In FTM HUCPVC cultures, cell surface MSC marker (CD90, CD105) expression and cell differentiation-related gene expression patterns were significantly more homogeneous in column-augmented vessels. MEF cells in column-augmented culture vessels showed a more consistent expression of IGF-1. Column-augmented cell culture vessels significantly improve the homogeneity of adherent cell cultures by mitigating the adverse effect of the secondary flow.This article has an associated First Person interview with the first author of the paper.
高质量的细胞培养需要可靠的实验室操作规范。如今，小规模细胞培养形式主要采用圆形拓扑结构的容器，每次重新定位细胞培养物时都会产生二次流，这是其固有的缺点。二次流会产生不均匀的沉降和黏附，对细胞培养质量产生负面影响。在此，我们展示了一种对圆形培养容器的改进方法，可消除这些干扰。在细胞培养孔中增加一个中心柱以减少二次流。将人癌细胞系（BeWo、JEG-3）、间充质干细胞[人脐带血管周围细胞（HUCPVC）]和小鼠胚胎成纤维细胞（MEF）分别培养在有中心柱的培养孔和常规培养孔中。人癌细胞培养物在有中心柱的容器中显示出均匀的细胞密度，且活细胞数量显著更多。在FTM HUCPVC培养物中，有中心柱的容器中细胞表面MSC标志物（CD90、CD105）的表达和细胞分化相关基因的表达模式明显更均匀。有中心柱的培养容器中的MEF细胞显示出更一致的IGF-Ⅰ表达。有中心柱的细胞培养容器通过减轻二次流的不利影响，显著提高了贴壁细胞培养的均匀性。本文对该论文的第一作者进行了相关的第一人称访谈。
