I am having a cell culture issue with my fibroblast adherent cells. Below are some of the cell issues that I am having. Hoping somebody can help me out in identifying the cause or the solution to this.
- Many cell clusters in flask after recovery and all through passaging.
- Cells grouping together and not spreading out like fibroblast cells should.
- Low cell counts.
- Poor growth.
- Cell clusters in 96 well assay plates.
- Assays failing because we are getting erratic results.
- Short cells (not elongated).
We have performed many tests to identify the cause of this issue, such as:
- Contamination (bacterial, fungal and mycoplasma
- Change in components (FBS, Penn Strep, Trypsin, HBSS, Basal Media)
- Flask/assay plate/stripette/conical tube material
If it somehow was the basal media - What component/mineral could affect the cells this much by either being at too high/low concentration? The basal media has all the essential materials already added. We only add the FBS and Penn Strep.
Below are images of the abnormal cells.
Thanks in advance,