Research Of Efficient And Economical Feeder Cells Preparation System

The conventional methodology (CM) for preparing feeder cells has some issues needed tobe optimized:(1) low yield with high cost;(2) fail to mitotically inactivate fully;(3) susceptibleto change the biological behaviors of MEFs;(4) manipulation time is inflexible. Therefore, it isnecessary to find new approaches that can be used not only for the production of feeder cells ona large scale in a single batch, but proliferation of the feeder cells are sufficiently suppressed aswell. To this end, we recently established a suspension-adherence methodology (SAM) and aCELLSPIN System methodology (CSSM) on the basis of and by optimization for CM.Firstly, to optimize the CM, we developed the SAM. By this way, it is more sufficient toinhibit cell mitosis and save time. Meanwhile, the bad state cells could be screened and we couldget the cells in good state. Thereby, we could improve the quality of feeder cells. We also didn’tneed to pay close attention to MEFs, and avoided worrying about missing the optimal treatmentopportunity for80%-90%confluent. So, it was more convenient than CM.Next, to increase the yield and reduce costs, we re-optimized the SAM, and used aCELLSPIN System (5-75RPM) to prepare the feeder cells. In the system, the cells were culturedin suspension during the entire process. Cells were more likely to contact with MMC, whichimproved the efficiency to inactivate the feeder cells. Moreover, suspension culture without anenzymatic digestion during harvest of the cells reduces the potential damages caused bytrypsinzation, and maintains the good state of the feeder cells as much as possible. The yield ofsingle spinner flask is several times or even hundreds times compared with10cm-dish. Thus,application of CELLSPIN System, we could harvest more cells in a short time, and increasefeeder cell yield. The CSSM is feasible for a large-scale production of hESCs and hiPSCs.