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Volume 2, Issue 1 (Spring 2004)
ASJ 2004, 2(1): 1-9 |
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Rahbar Pour T, Saleh Nia M, Soleimani M. Differentiation of Mouse Yolk Sac Cells to Erythroid Cells in The Presence of Erythropoietin. ASJ 2004; 2 (1) :1-9
URL: http://anatomyjournal.ir/article-1-353-en.html
URL: http://anatomyjournal.ir/article-1-353-en.html
1- Anatomy Department, Tarbiat Modarres University, Tehran, Iran.
Abstract: (754 Views)
Purpose: Yolk sac hematopoietic stem cells (YS-HSC) have two dominant characteristices: a larger reproductive capacity and the absence of the expression of MHC associated antigens. Therefore, these cells are promising candidates for transplantation, cell therapy and gene manipulation. There are controversial reports on the effects of erythropoietin (EPO) on the differentiation of yolk sac cells (YSC) to erythroid lineage in vitro.
Materials and Methods: The yolk sacs were dissected from 10 - day - old NMRI mouse embryos and separated mechanically and enzyme digestions (0.1% collagenase or 0.25% trypsin/O.02% EDTA). The colony assay of these cells was performed in the presence of interleukin-3, stem cell factors and different dosages of EPO in a semisolid medium. A control group was also considered in the absence of EOP. After 14 days, benzidine staining on the colonies was carried out and positive benzidine colonies were counted as erythroid colonies. Also, the cytospine slides of those colonies were prepared and stained using benzidine and wright giemsa staining. There was not good result from direct culturing of yolk sac cells. So these cells were co-cultured on the bone marrow stromal cells as a feeder layer.
Results: The results showed that in the presence and absence of EPO, the YSCs were differentiated to erythroid cells. However, the number of the positive benzidine colonies (55.4%) and the size of colonies in the presence of EPO (1 U/ml) were increased compared to the control group. There was no statistically significant difference between these groups. Co-culture of these cells with feeder layer also revealed that in the presence of EPO, a better differentiating effect could be observed.
Conclusion: In conclusion, in vitro differentiation of YSCs to erythroid lineage was seen in the absence of EPO. However EPO could have a positive effect on the improvement of the erythroid colonies, it needs more investigation.
Materials and Methods: The yolk sacs were dissected from 10 - day - old NMRI mouse embryos and separated mechanically and enzyme digestions (0.1% collagenase or 0.25% trypsin/O.02% EDTA). The colony assay of these cells was performed in the presence of interleukin-3, stem cell factors and different dosages of EPO in a semisolid medium. A control group was also considered in the absence of EOP. After 14 days, benzidine staining on the colonies was carried out and positive benzidine colonies were counted as erythroid colonies. Also, the cytospine slides of those colonies were prepared and stained using benzidine and wright giemsa staining. There was not good result from direct culturing of yolk sac cells. So these cells were co-cultured on the bone marrow stromal cells as a feeder layer.
Results: The results showed that in the presence and absence of EPO, the YSCs were differentiated to erythroid cells. However, the number of the positive benzidine colonies (55.4%) and the size of colonies in the presence of EPO (1 U/ml) were increased compared to the control group. There was no statistically significant difference between these groups. Co-culture of these cells with feeder layer also revealed that in the presence of EPO, a better differentiating effect could be observed.
Conclusion: In conclusion, in vitro differentiation of YSCs to erythroid lineage was seen in the absence of EPO. However EPO could have a positive effect on the improvement of the erythroid colonies, it needs more investigation.
Type of Study: Original |
Subject:
Morphometry
Received: 2021/12/25 | Accepted: 2004/04/22 | Published: 2004/04/22
Received: 2021/12/25 | Accepted: 2004/04/22 | Published: 2004/04/22
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Anatomical Sciences Journal (ASJ)
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