I am using these product from ATCC (Primary Uterine Fibroblast Cells; Normal, Human (HUF) ATCC Cat. No. PCS-460-010 ) with their recommended media and with Growth Kit-Low Serum aacording to their recommendation. But I found the expression of alpha SMA, FAP and other CAF marker even without activation by cancer cell conditioned media. Please suggest me what to do ?
Possibly the protein which might be activated is absent or could not be activated since it was linked to another compound or protein
Hello Subhadip Kundu
I feel these normal fibroblasts are getting activated during invitro propagation. There is a possibility that the culture conditions may be contributing to the activation of these normal fibroblasts. These fibroblasts can get activated in response to various stimuli present in the culture environment. One such stimuli being the growth factors present in the culture media or some kind of stress experienced by the cells in culture.
So, try to find the source and adopt measures to minimize the activation of these cells.
Best.
Begin by testing for mycoplasma contamination (this happens even in big pharma). The choice of plasticware is also crucial for these primary cells, so ensure strict adherence to the established SOPs for both maintenance and cell detachment. If problems persist despite following the correct procedures, consider requesting a new vial (for free) from ATCC. Good luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue