[In vitro study on the transduction of mutant human dihydrofolate reductase cDNA into human umbilical cord blood CD34+ cells].
OBJECTIVE
To explore whether human umbilical cord blood hematopoietic progenitor cells transduced with mutant dihydrofolate reductase(mDHFR) gene increase resistance to metrotrexate(MTX).


METHODS
Cord blood CD34+ cells were enriched with a high-gradient magnetic cell sorting system(MACS), and then transfected with supernatant of retrovirus containing human mDHFR cDNA. Hematopoietic progenitor clonogenic assay was used to evaluate mDHFR-conferred resistance to MTX.


RESULTS
The purity of cord blood CD34+ cells was approximately 90% and recovery rate was 71.1%. The transduced and mock-transduced CD34+ cells were cultured in mediums containing 20 nmol/L MTX for 14 days. The percentages of progenitor colonies of transduced and mock-transduced CD34+ cells were higher than that of non-transduced (P < 0.01), obviously. The resistance of the former to MTX is nearly twofold of the latter.


CONCLUSIONS
Transduction of mDHFR gene could confer the resistance of human hematopoietic progenitor cells to MTX toxicity.
Wu
R
R
Cancer Institute, CAMS and PUMC, Beijing 100021.
Yang
Z
Z
Liu
J
J
D018952:Antigens, CD34 / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008727:Methotrexate / Q000494:pharmacology; D009154:Mutation; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014162:Transfection
D018952:Antigens, CD34; D018076:DNA, Complementary; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate