In vivo expression of human growth hormone by genetically modified murine bone marrow stromal cells and its effect on the cells in vitro.
Human growth hormone (hGH) is frequently used clinically for growth abnormalities in children and also in adults with growth hormone deficiency. The hormone is usually administered to the individuals by frequent injections. In the present study we investigated the potential of bone marrow stromal cells as vehicles to deliver the GH in vivo by infusion of cells transduced with hGH cDNA into mice femurs. The effect of the hormone on the transduced cells in vitro was also assessed. Bone marrow stromal cells established from a mouse model of human osteogenesis imperfecta mice (oim) were transduced with a retrovirus containing hGH and neomycin resistance genes. The hGH-expressing cells were selected in a medium containing G418 and were then assessed for the hGH expression in vitro. The selected cells synthesized 15 ng/10(6) cells of hGH per 24 h in vitro and exhibited alkaline phosphatase activity when they were treated with the human recombinant bone morphogenetic protein 2 (rhBMP-2). The transduced cells also proliferated faster than the LacZ transduced cells but they did not exhibit a higher rate of matrix synthesis. When 2 x 10(6) hGH+ cells were injected into the femurs of mice, hGH was detected in the serum of the recipient mice up to 10 days after injection. The highest level of growth hormone expression, 750 pg/ml, was detected in the serum of the recipient mice I day after injection of the transduced cells. hGH was also detected in the medium conditioned by cells that were flushed from the femurs of the recipient mice at 1, 3, and 6 days after cell injection. These data indicate that bone marrow stromal cells could potentially be used therapeutically for the delivery of GH or any other therapeutic proteins targeted for bone. The data also suggest that GH may exert its effects on bone marrow stromal cells by increasing their rate of proliferation.
Suzuki
K
K
Musculoskeletal Research Center, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, PA 15261, USA.
Oyama
M
M
Faulcon
L
L
Robbins
P D
PD
Niyibizi
C
C
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology*; D016026:Bone Marrow Transplantation; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000494:pharmacology; D002455:Cell Division; D048429:Cell Size; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D015316:Genetic Therapy / Q000379:methods*; D019382:Human Growth Hormone / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D006801:Humans; D007763:Lac Operon / Q000235:genetics; D051379:Mice; D010013:Osteogenesis Imperfecta; D011994:Recombinant Proteins / Q000494:pharmacology; D017154:Stromal Cells / Q000502:physiology* / Q000637:transplantation; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D019382:Human Growth Hormone; D003094:Collagen; D000469:Alkaline Phosphatase