Hematopoietic Recovery after Transplantation CD117+B220- (lacZ+) Bone Marrow Cells in Lethally Irradiated Mice

Miroslav Hodek,Stanislav Filip,Zuzana Šinkorová,Jaroslav Mokrý,Jiřina Vávrová

doi:10.14712/18059694.2017.6

Abstract

Experiments presented here were aimed at the description of hematopoiesis repair and in vivo homing of transplanted separated CD117+B220- bone marrow cells after whole-body lethal irradiation at LD 9Gy. ROSA 26 mice were used as donors of marrow cells for transplantation [B6;129S/Gt (ROSA)26Sor] and were tagged with lacZ gene, and F2 hybrid mice [B6129SF2/J] were used as recipients of bone marrow transplanted cells. Hematopoiesis repair was provided by transplantation, both suspension of whole bone marrow cells (5x106) and isolated CD117+B220- cells (5x10(4)). Mice survived up to thirty days after irradiation. We demonstrated that transplantation of suspension of whole bone marrow cells led to faster recovery of CFU-GM (Granulocyte-macrophage colony forming units) in bone marrow and in the spleen too. It is not clear what the share of residential and transplanted cells is in the repair process. Our results demonstrate that sufficient hematopoietic repair occurs after transplantation of CD117+B220- (lacZ+) in lethally irradiated mice, and the difference in CFU-GM numbers in the bone marrow and spleen found on day 8 posttransplant has no influence on the survival of lethally irradiated mice (30 days follow-up).

Highlights

Hematopoietic stem cells (HSCs) in mice are defined as having self-renewal ability, and can repair all blood cell lineages
In our experiments we studied the regeneration of hematopoiesis in lethally irradiated mice after transplantation of separated CD117+ marrow cells and the relationship between kinetics and in vivo homing in resident CD117+ cells and transplanted CD117+ cells
Hematopoietic rescue was noted through culture of CFU-GM in the bone marrow (Fig. 2) and spleen in lethally whole-body irradiated mice exposed to LD 9 Gy

Summary

Introduction

Hematopoietic stem cells (HSCs) in mice are defined as having self-renewal ability, and can repair all blood cell lineages. Another example may be of bone marrow cells in a mouse population with phenotypic characteristic of resting HSCs but without renewing hematopoiesis in irradiated mice These cells express high levels of Sca-1, H-2K and CD38 and low levels of Thy-1.1 but did not express CD34 or lineage markers CD3, CD4, CD5, CD8, NK1.1, I-A, B220, Ig(MGA), CD40, Mac-1, Gr-1 and Ter119. The phenotypically characterized cells of Sca-1+, Thy-1.1low (cell surface markers) and their insufficient expression of c-kit do not respond adequately to hematopoietic growth factors in vitro. They do not form spleen colonies and have no ability to reconstitute hematopoiesis in irradiated mice [12]

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Discussion

Conclusion