I wanted to create feeder layers of chicken embryo fibroblasts and testis somatic cells for growing of chicken spermatogonial stem cells.
When I grew the cells up to full confluent all the layer detached from the plate soon after mitimycine C treatment. Then I treated them at 80% confluency, but when I add there the suspension of testis cells containing as well as spermatogonial stem cells and somatic cells they grow rapidly and in a day or two also start to detach from the plate. If I passage them so frequently Im afraid to loose all the SSCs.
I would highly appreciate if anybody prompt how to deal the problem