American Society of Hematology

Fig. 2.

$Fig. 2. Limiting-dilution transplant analysis of MDR1-transduced SP cells isolated from 12-day expansion cultures. / HaMDR1-transduced BM cells (C57BL/6 background) were placed in liquid suspension culture and expanded for 12 days. On day 12, the SP cell fraction was isolated by flow cytometry using the sorting gate shown in A. The sorted cells were then injected into lethally irradiated recipient mice at the indicated doses, along with a 2 × 105 fresh BM cells (HW80 background). (A) 16 weeks after transplantation, peripheral blood leukocyte DNA was prepared and analyzed by PCR for the presence of the HaMDR1 proviral genome. A water-only control and a nontransplanted mouse control (HW80) are shown on the left as negative controls. Transplanted mice received cell doses ranging from 250 to 9400 cells, as indicated above the lanes. The numbers 1 to 18 correspond to individual recipient mice. (B) Reconstitution was also measured using hemoglobin electrophoresis. Erythroid cells derived from the sorted SP cells are identified by the faster-migrating, C57BL/6-derived hemoglobin isoform. The animal numbers are the same as in A, and samples from nontransplanted C57BL/6 and HW80 mice are shown on the right. Asterisks indicate samples in which there were detectable amounts of C57BL/6-derived hemoglobin.$

Limiting-dilution transplant analysis of MDR1-transduced SP cells isolated from 12-day expansion cultures.

HaMDR1-transduced BM cells (C57BL/6 background) were placed in liquid suspension culture and expanded for 12 days. On day 12, the SP cell fraction was isolated by flow cytometry using the sorting gate shown in A. The sorted cells were then injected into lethally irradiated recipient mice at the indicated doses, along with a 2 × 10⁵ fresh BM cells (HW80 background). (A) 16 weeks after transplantation, peripheral blood leukocyte DNA was prepared and analyzed by PCR for the presence of the HaMDR1 proviral genome. A water-only control and a nontransplanted mouse control (HW80) are shown on the left as negative controls. Transplanted mice received cell doses ranging from 250 to 9400 cells, as indicated above the lanes. The numbers 1 to 18 correspond to individual recipient mice. (B) Reconstitution was also measured using hemoglobin electrophoresis. Erythroid cells derived from the sorted SP cells are identified by the faster-migrating, C57BL/6-derived hemoglobin isoform. The animal numbers are the same as in A, and samples from nontransplanted C57BL/6 and HW80 mice are shown on the right. Asterisks indicate samples in which there were detectable amounts of C57BL/6-derived hemoglobin.

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