The Effect of Cryopreservation of Bone Marrow Cells from Donor Mice that Carry the egfp Gene, on the Lifespan of Mice after Syngeneic Transplantation

Abstract

The effect of cryopreservation of bone marrow cells on the lifespan of mice after syngeneic transplantation has been studied in nonirradiated mice and 7 Gy-irradiated mice. Mice with the enhanced green fluorescent protein gene were the donors. Bone marrow cells were cryopreserved according to the method used in clinical practice in the field of bone marrow autotransplantation in the treatment for patients with cancer. Dimethyl sulfoxide dissolved in polyglucin at the final concentration of 5% acted as a cryoprotectant agent. Transplantation of the thawed stem cells was carried out without washing out the cryoprotectant. No side effects associated with the cryoprotectant toxicity were observed. It has been shown that staining of bonemarrow cells with trypan blue is a more selective technique to evaluate the extent of cell damage after cryopreservation. The mean lifespan of nonirradiated recipient mice was not statistically different from that of the intact control group. In irradiated recipient mice, the mean lifespan increased by 51 ± 2% compared to the group of irradiated controls. The analysis of a blood sample taken from the tail vein of irradiated mice revealed lifelong engraftment of donor-derived cells in the hematopoietic system of the recipient mice. Thus, model experiments on the syngeneic strain of mice showed that cryopreserved bone-marrow cells can be effectively used for cell therapy in autotransplantation in patients after X-ray radiation therapy.