Document Type : Original Article
Introduction: Setting up a system to study the in vitro hematopoiesis is helpful to understand the mechanism of hematopoiesis. The effects of several cytokines and hormones such as erythropoietin (EPO) in differentiation of hematopoietic stem cells have been investigated in vivo. There are no records considering the potential differentiation of P19 cell line (embryonic carcinoma cells) to hematopoietic cells. The aim of this study was to use this cell line as a model to evaluate the effects of interleukin 3, 6 and erythropoietin on the differentiation of stem cells.
Material and Methods: P19 cells were cultured directly on semisolid medium supplemented with 10% fetal calf serum (FCS). The embryoid bodies were formed 9 days after culturing. The cells were trypsinized and dissociated to single cells. These cells were cultured on the semisolid medium containing 10% FCS, 10 ng/ml IL-3 and IL-6 and in the presence and absence of 20 u/ml erythropoietin. The colonies were formed after 14 days. Then the colonies were studied by benzidine and giemsa staining.
Results: Our results showed that in the presence of IL-3, IL-6 and EPO the positive and negative benzidine colonies were 54% and 46%, respectively. In the absence of EPO the mentioned colonies were 14% and 86%, respectively. There was a statistically significant difference between the positive and negative colonies in both groups (p<0.001).
Conclusion: Under the effects of IL-3, IL-6 and EPO, the P19 cells could differentiate to erythroid and non-erythroid lineage and in the presences of EPO the umber of erythroid colonies increased.