embryos (13-14 days of gestation)
were decapacitated and it was minced finely in the
dish containing Trypsin/EDTA then the tissue was
incubated for 30 minutes. The tissue was transferred
into 50 ml cornical tube and the supernatant
was pipetted into a new tube, cells were pelleted
and seeded on 25 cm2 culture flask (Nunc) in αMEM
supplemented with 10% FCS. At subconfluence,
fibroblasts were trypsinized and cultured up to the
third passage and frozen the same as donor cells.
Frozen-thawed fibroblasts were cultured in αMEM
supplemented with 10% FCS and used as co-culture
cell by culture in SOFaa-10 4 days before use
at concentration 106 cell/ml.