Engraftment of stem-cell-enriched bone marrow fractions in MHC-identical dogs after fractionated total-body irradiation

Abstract

Discontinuous albumin density gradients were used to obtain enrichment of hemopoietic stem cells and depletion of T lymphocytes in aspirated dog bone marrow. Colony forming units in agar (CFU-C) were determined to evaluate the degree of enrichment achieved. An average CFU-C concentration factor of 12.4 was obtained. All transplantations in the study were carried out between DLA-identical sibling combinations. The number of CFU-C administered varied from 0.2 to 5.5 X 10(5)/kg and the number of nucleated cells transfused varied from 0.1 to 1.0 X 10(8)/kg. Stem cell concentrates were found more difficult to engraft than unmodified bone marrow following standard conditioning with a single total-body irradiation (TBI) dose of 7.5 Gy. The efficacy of different TBI-fractionation schedules for obtaining sustained engraftment of CFU-C-enriched grafts in identical bone marrow transplantation (BMT) was determined. A total dose of 12 Gy TBI delivered in two equal fractions of 6.0 Gy (72-hr interval) resulted in sustained engraftment of stem cell grafts in 7 of 7 evaluable dogs. A TBI dose of 9 Gy in two fractions of 4.5 Gy (72-hr interval) resulted in sustained engraftment in 5 of 7 evaluable dogs. The two dogs with engraftment failure received low total cell numbers (10(7) cells/kg) and low CFU-C numbers. 9 Gy of TBI in two fractions of 4.5 Gy (24-hr interval) resulted in sustained engraftment in 11 of 12 evaluable dogs. A significant improvement of engraftment was obtained by increasing the total dose of TBI, which necessitates fractionation into two fractions of TBI. The lower-total-dose TBI (9 Gy) produced less early and late toxicity than the total high-dose (12 Gy) TBI. The incidence of engraftment was similar for the two dosages, however the recovery of peripheral leukocyte counts was slower after 9 Gy TBI. In the dog, optimal conditioning for lymphocyte-depleted hemopoietic stem cell grafts can be obtained by increasing the dose of TBI and concomitant fractionation.