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J Biosci Bioeng ; 124(5): 591-598, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28688754

RESUMO

If cultured in appropriate conditions, such as supplementing culture media with costly cytokines and growth factors, hematopoietic stem/progenitor cells (HSPCs) from different origins have shown to be an adequate source of erythroid cells. This requirement turns erythroid cells production into a complicated process to be scaled-up for future applications. The aim of our work was to genetically modify HSPCs with human erythropoietin (hEPO) sequence by lentiviral transgenesis in order for cells to secrete the hormone into the culture medium. Initially, we evaluated erythroid differentiation in colony forming units (CFU) assays and further analyzed cell expansion and erythroid differentiation throughout time in suspension cultures by flow cytometry and May-Grünwald-Giemsa staining. Additionally, we studied hEPO production and its isoforms profile. The different assessment approaches demonstrated erythroid differentiation, which was attributed to the hEPO secreted by the HSPCs. Our data demonstrate that it is possible to develop culture systems in which recombinant HSPCs are self-suppliers of hEPO. This feature makes our strategy attractive to be applied in biotechnological production processes of erythroid cells that are currently under development.


Assuntos
Técnicas de Cultura de Células/métodos , Diferenciação Celular/genética , Células Eritroides/citologia , Eritropoetina/genética , Eritropoetina/metabolismo , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Lentivirus/genética , Biotecnologia/métodos , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Células Eritroides/metabolismo , Eritropoetina/biossíntese , Eritropoetina/química , Humanos , Lentivirus/metabolismo
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