When working with primary cells, it is important to remember that they are not cell lines and cannot be treated the same. Because iXCells Biotechnologies has extensively worked with primary cells, we are very familiar with the common problems researchers encounter when culturing them. Here are five common mistakes that are made with primary cells and how to correct these mistakes.
Correction #1: Primary cells are very sensitive to the thawing process so it is important that the vial be placed in a 37oC water bath, held and rotated gently until the contents are just thawed. Then remove the vial from the water bath promptly and transfer in into a sterile hood. Make sure your culture vessel is ready before thawing so the cells can immediately be seeded and placed in the incubator.
Correction #2: Primary cells can become senescent when grown to 100% confluent. Remember that primary cells are not 100% pure, so it is important to minimize growth of contaminating cells. We recommend subculturing primary cells when they are 90-95% confluent.
Correction #3: When passaging cells, use low concentrations of trypsin and monitor cells closely under the microscope. Also, remember to completely neutralize the cells after trypsinization because any active trypsin will damage the cells.
Correction #4: Normally we do not recommend refreezing primary cells as this can promote a senescent phenotype and/or cause functional changes. Primary cells are extremely sensitive and refreezing may result in cell death or damage.
Correction #5: Unlike cell lines, primary cells have a limited expansion capacity. We recommend using primary cells as early as possible for experiments to prevent genetic drift. In addition, if you are working with a difficult cell type you should monitor cell morphology closely for contaminating cells as they can take over the culture over time.
Access Dr. Nianwei Lin’s presentation to gain valuable scientific insights
luanamswilliams
Multi-electrode array recordings of spontaneous activity of glutamatergic neurons for
luanamswilliams
Peripheral Blood Mononuclear Cells (PBMCs) is any blood cell with
luanamswilliams
iXCells Biotechnologies is proud to provide the world’s first commercial,
luanamswilliams
Adipose Tissue Derived Cells Overview Adipose tissue, or body fat,
luanamswilliams
immune AND hematopoietic cells Your dependable source of human primary
luanamswilliams
Proin sit amet urna nec arcu consequat luctus sed id
luanamswilliams
