Low level of gene transfer to and engraftment of murine bone marrow cells from long-term bone marrow cultures
| Autor: | Thomas Relander, Cecilia Fahlman, Stefan Karlsson, Johan Richter |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
medicine.medical_specialty Transplantation Conditioning Cell Bone Marrow Cells Biology Transfection Time Green fluorescent protein Mice Mice Congenic Internal medicine Genetics medicine Animals Protamines Progenitor cell Molecular Biology Cells Cultured Bone Marrow Transplantation Stem Cell Factor Hematology Interleukin-6 Graft Survival Gene Transfer Techniques Heparin Antagonists Cell Biology Flow Cytometry Molecular biology Coculture Techniques Culture Media Mice Inbred C57BL Survival Rate Haematopoiesis medicine.anatomical_structure Cell culture Bone marrow Stem cell Cell Division |
| Zdroj: | Experimental Hematology. 28:373-381 |
| ISSN: | 0301-472X |
| DOI: | 10.1016/s0301-472x(00)00131-4 |
| Popis: | OBJECTIVE: We wanted to determine whether the long-term bone marrow culture (LTBMC) transduction system would lead to efficient gene transfer and engraftment of murine repopulating hematopoietic stem cells (HSC), particularly in nonablated recipients. MATERIALS AND METHODS: Congenic mouse strains expressing Ly 5.1 or Ly 5.2 and the GP+E86 cell line producing the MGirL22Y vector carrying the gene for enhanced GFP were used. Murine LTBMCs were established and demi-depopulated on days 7 and 14 with addition of vector supernatant on days 8 and 15. RESULTS: Cell recovery on day 21 was 21.3%+/-3.8% of input cells and CFU-C recovery was 9.7+/-3.4% as compared with CFU-C of input cells. In vitro transduction efficiency determined by CFU-C expressing GFP was 22.2%+/-1.6%. In irradiated (950 cGy) mice transplanted with 2x10(6) LTBMC cells, 94% of nucleated cells in the blood at week 16 were of donor origin. However, GFP was only detected at low level in a few animals at week 4 and not later. Analysis of bone marrow from these mice at week 20 did not show any GFP expression and semiquantitative PCR revealed a transgene level of |
| Databáze: | OpenAIRE |
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