High-level expression of mouse growth hormone (mGH) by primary human keratinocytes using a retroviral vector
Carregando...
Data
Data de publicação:
2002
Orientador
Título da Revista
ISSN da Revista
Título do Volume
É parte de
É parte de
É parte de
É parte de
REUNIAO ANUAL DA SOCIEDADE BRASILEIRA DE BIOQUIMICA E BIOLOGIA MOLECULAR, 31.
Exportar
Resumo
The skin is considered a potential target for the development of gene therapy protocols for cutaneous or systemic diseases. The high proliferative capacity of cultured keratinocytes makes these epidermal colls idoal candidates for genetie manipulation. We have utilized primary human keratinocytes transduced with an efficient retroviral vector (LXSN) encoding a mouse growth hormone gene (mGH). The main goals are the conetruction of an homologous ox vivo transfer protocol and the comparison with a similar however heterologous system based on the human growth hormone (NGH) gene. After transfection of the vector LmGHSN into GP + E-86 ecotropic packaging cell lins, tho exprassion of mGH (10ng/mL) was detected in the culture medium by a specific radioimmonuassay. This supernatant was then used to infect the amphotropic packaging cell lins GP + env Am12 and 36 clones were isolated by G418 selection. Clones presenting a secretion lovel ranging from 22-50ng mGH/10(sup)6 cells.day (44 to 150ng mGH/mL) and a viral titer of 10(sup)5 - 10(sup)6 CFU/mL were chosen to infect the keratinocytes. The modified keratinocytes presented a stable in vitro secretion level up to 3ug mGH/10(sup)6cells.day (approximately 4ug mGH/mL), during serial propagation. These results are analogous to those previously obtained for the HGH model, which was already utilized for grafting into immunodeficient dwarf mice (little/scid), Halt-ifo dotorminations for hGH and mGH are now being carried out trying to verify a possible difference of plasma clearance rate in mice, in order to correlats these data with the lvels of GH socreted in vivo by the gonotically modified keratinocytes.
Como referenciar
ROSAURO, C.W.; PERONI, C.N.; POIATO, E.E.; SOUSA, J.M.; BARTOLINI, P. High-level expression of mouse growth hormone (mGH) by primary human keratinocytes using a retroviral vector. In: REUNIAO ANUAL DA SOCIEDADE BRASILEIRA DE BIOQUIMICA E BIOLOGIA MOLECULAR, 31., 18-21 maio, 2002, Caxambu, MG. Resumos... p. 106. Disponível em: http://repositorio.ipen.br/handle/123456789/20940. Acesso em: 20 Mar 2025.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.