Dear authors,
I have followed this and similar procedures to try to isolate mouse fibroblasts but none of them are working as I'm not getting many fibroblasts to adhere to the cell culture flask. I'm using minced tissue from the hearts of 3 adult mice and after a couple hours, there are small patches of healthy fibroblasts but not even enough to be 50% confluent in one well of a 12 well plate.
I'm either killing the healthy fibroblasts or something is preventing them from adhering to the plate. I've done everything in this procedure as stated so I'm having a lot of trouble pinpointing where exactly things go wrong. I've also repeated similar procedure from published literature (very similar to this one) again with marginal yield.
What are the most error-prone steps in this procedure/where are the places where I should start in troubleshooting?
Thanks in advance.
-T Blake Monroe
5/29/2018 8:15:28 AM Reply