[Reconstitution and protection of murine hematopoiesis by second transplantation with bone marrow cells containing mutant human DHFR gene from primary generation murine].
OBJECTIVE
To observe the effect of donor murine bone marrow, which was transfected in vivo with mutant hDHFR gene, on the reconstitution and protection of hematopoiesis of lethally irradiated recipient mice.


METHODS
Mice bone marrow transduced in vivo with mutant hDHFR gene were used for transplantation. After whole bodily irradiated (8.5-9.0 Gy) and then transplanted with the transduced marrow, the recipients were treated with methotrexate (4 mg/kg twice a week for the first week; and then 10 mg/kg twice a week). The survival rate and blood picture of the recipients as well as the integration and expression of hDHFR gene were studied by using PCR and Southern blotting.


RESULTS
The hematopoiesis of the recipient mice could be reconstituted and protected from lethal toxicity of high dose methotrexate. It was confirmed that hDHFR gene was integrated into hematopoietic cell as well as some tissues genomic DNA and expressed successfully in the recipients.


CONCLUSIONS
Donor mice bone marrow transfected with mutant hDHFR gene can effectively reconstitute and protect hematopoiesis in lethally irradiated recipient mice.
Zhang
W
W
Beijing Institute for Cancer Research, School of Oncology, Beijing Medical University, Beijing 100034.
Xu
Z
Z
Xu
G
G
D000818:Animals; D001853:Bone Marrow / Q000378:metabolism*; D016026:Bone Marrow Transplantation / Q000379:methods*; D006410:Hematopoiesis / Q000502:physiology*; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D011897:Random Allocation; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection; D014916:Whole-Body Irradiation
C113155:DHFR-ASC; D013762:Tetrahydrofolate Dehydrogenase