Stimulation of retroviral vector infection of murine hematopoietic progenitors.
Conditioned media (CM) from a cloned murine marrow-derived stromal cell line, AC6.21 (ALC), was shown to stimulate retroviral vector infection of hematopoietic progenitors in culture. Inclusion of ALC CM during cocultivation of normal murine bone marrow (BM) with vector-producing fibroblasts improved infection efficiency of day 13 spleen colony-forming cells (CFU-s) from 63% (15 provirus-positive spleen colonies/24 total), without added growth factor, to 90% (36 provirus-positive colonies/40 total). In addition, stimulation of BM cells with ALC CM during cocultivation improved retroviral infection of stem cells capable of repopulating the hematopoietic system of irradiated recipient animals. Because ALC CM was found to have 50 to 100 U/ml of IL-6 activity, purified recombinant human IL-6 was tested for an effect in this system. Stimulation with IL-6 alone increased retroviral infection efficiency of CFU-s from 15% (17 colonies provirus-positive/111 total analyzed) without added growth factor to 66% (97 provirus-positive colonies/148 total analyzed). These experiments support and extend previous studies which have demonstrated the necessity for growth factor stimulation in optimizing retroviral vector transduction of hematopoietic precursors.
Fletcher
F A
FA
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Belmont
J W
JW
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003114:Colony-Forming Units Assay; D003470:Culture Media; D004279:DNA, Viral; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000382:microbiology*; D007377:Interleukin-3 / Q000235:genetics / Q000494:pharmacology; D015850:Interleukin-6 / Q000235:genetics / Q000494:pharmacology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D003470:Culture Media; D004279:DNA, Viral; D007377:Interleukin-3; D015850:Interleukin-6; D011994:Recombinant Proteins