6 common mistakes in primary cell culture
2016-10-17 Beijing Yuhengfeng Technology
When culturing primary cells, it is important to remember that they are not cell lines and cannot be treated the same. Because of the extensive work of ScienCell in primary cell culture, we are very familiar with the common problems that researchers encounter when culturing primary cells. Here are six common mistakes in primary cell culture and how to correct them.
Error #1: A small tube of primary cells is thawed in a water bath for a while
Correction #1: Primary cells are very sensitive to the thawing process, so it is important to place the vial in a 37 ° C water bath, hold and gently rotate until the contents have just thawed. The vial should then be removed from the water bath immediately and transferred to the aseptic workstation. Make sure your flask is ready before thawing so that it can be used immediately to seed the cells and placed in the incubator.
Error #2: Centrifuge the primary cells directly after thawing the vial
Correction #2: We do not recommend centrifuging the cells after thawing because the centrifugation procedure is more harmful than a small amount of DMSO residue. Remember to change the medium the next day after restoring the primary cells to remove any residual DMSO.
Error #3: Allow primary cells to become too fused
Correction #3: Primary cells can become senescent when grown to 100% confluence. Remember that primary cells are not 100% pure, so it is important to minimize the growth of contaminating cells. We recommend subculture of primary cells when the cells reach 90-95% confluency.
Error #4: Excessive trypsin digestion when passaged primary cells
Correction #4: When cells were passaged, low concentrations of trypsin were used and the cells were closely monitored under a microscope. In addition, remember to completely neutralize trypsin in the cells after trypsinization, as any active trypsin will damage the cells. We specifically recommend a trypsin neutralization solution (Cat#0113) specifically formulated for primary cells, but 10% serum or soybean trypsin inhibitor (Cat#0173) can also be used.
Error #5: Primary cells can be easily re-frozen
Correction #5: Usually we do not recommend re-frozen primary cells as this can promote cellular senescence and/or cause functional changes. Primary cells are very sensitive and re-frozen can cause cell death or damage.
Mistake 6: Primary cells can proliferate indefinitely
Correction #6: Unlike cell lines, primary cells have limited amplification capabilities. We recommend using primary cells as early as possible to experiment to prevent genetic drift. In addition, if you are using a cell type that is difficult to add value, you should closely monitor the cell morphology, as a small number of mixed cells (such as fibroblasts) may grow in large quantities over time to become the main cell.
It is important to keep these six technical tips in mind when culturing primary cells. Ultimately, if you treat your cells carefully and carefully, they will better serve your experiment.
If you have any other technical questions about primary cell culture, you can contact Yuhengfeng Technology's technical support.
Yu Hengfeng (America's Sciencell agent) order hotline, 63803739, 63861816, 63822556
For more cell consultation, please contact us.
Organic Garlic Flakes,Organic Roasted Garlic Flakes,Organic Garlic Flakes For Sale,Organic Dried Garlic Flakes
shandong changrong international trade co.,ltd. , https://www.changronggarliccn.com