Plant-produced human recombinant erythropoietic growth factors support erythroid differentiation in vitro.

Musiychuk, Konstantin; Sivalenka, Rajarajeswari; Jaje, Jennifer; Bi, Hong; Flores, Rosemary; Shaw, Brenden; Jones, R Mark; Golovina, Tatiana; Schnipper, Jacob; Khandker, Luipa; Sun, Ruiqiang; Li, Chang; Kang, Lin; Voskinarian-Berse, Vanessa; Zhang, Xiaokui; Streatfield, Stephen; Hambor, John; Abbot, Stewart; Yusibov, Vidadi

Musiychuk, Konstantin; Sivalenka, Rajarajeswari; Jaje, Jennifer; Bi, Hong; Flores, Rosemary; Shaw, Brenden; Jones, R Mark; Golovina, Tatiana; Schnipper, Jacob; Khandker, Luipa; Sun, Ruiqiang; Li, Chang; Kang, Lin; Voskinarian-Berse, Vanessa; Zhang, Xiaokui; Streatfield, Stephen; Hambor, John; Abbot, Stewart; Yusibov, Vidadi.

Afiliación

Musiychuk K; Fraunhofer USA Center for Molecular Biotechnology, Newark, DE 19711, USA.

Stem Cells Dev ; 22(16): 2326-40, 2013 Aug 15.

Article en En | MEDLINE | ID: mdl-23517237

RESUMEN

Clinically available red blood cells (RBCs) for transfusions are at high demand, but in vitro generation of RBCs from hematopoietic stem cells requires significant quantities of growth factors. Here, we describe the production of four human growth factors: erythropoietin (EPO), stem cell factor (SCF), interleukin 3 (IL-3), and insulin-like growth factor-1 (IGF-1), either as non-fused proteins or as fusions with a carrier molecule (lichenase), in plants, using a Tobacco mosaic virus vector-based transient expression system. All growth factors were purified and their identity was confirmed by western blotting and peptide mapping. The potency of these plant-produced cytokines was assessed using TF1 cell (responsive to EPO, IL-3 and SCF) or MCF-7 cell (responsive to IGF-1) proliferation assays. The biological activity estimated here for the cytokines produced in plants was slightly lower or within the range cited in commercial sources and published literature. By comparing EC50 values of plant-produced cytokines with standards, we have demonstrated that all four plant-produced growth factors stimulated the expansion of umbilical cord blood-derived CD34+ cells and their differentiation toward erythropoietic precursors with the same potency as commercially available growth factors. To the best of our knowledge, this is the first report on the generation of all key bioactive cytokines required for the erythroid development in a cost-effective manner using a plant-based expression system.

Asunto(s)

Diferenciación Celular/efectos de los fármacos; Eritropoyetina/farmacología; Células Madre Hematopoyéticas/efectos de los fármacos; Factor I del Crecimiento Similar a la Insulina/farmacología; Interleucina-3/farmacología; Nicotiana/genética; Factor de Células Madre/farmacología; Agrobacterium tumefaciens/genética; Línea Celular Tumoral; Proliferación Celular/efectos de los fármacos; Clonación Molecular/métodos; Eritrocitos/citología; Eritrocitos/metabolismo; Eritropoyetina/biosíntesis; Eritropoyetina/genética; Eritropoyetina/aislamiento & purificación; Sangre Fetal/citología; Sangre Fetal/efectos de los fármacos; Sangre Fetal/metabolismo; Expresión Génica; Vectores Genéticos; Células Madre Hematopoyéticas/citología; Células Madre Hematopoyéticas/metabolismo; Humanos; Factor I del Crecimiento Similar a la Insulina/biosíntesis; Factor I del Crecimiento Similar a la Insulina/genética; Factor I del Crecimiento Similar a la Insulina/aislamiento & purificación; Interleucina-3/biosíntesis; Interleucina-3/genética; Interleucina-3/aislamiento & purificación; Plantas Modificadas Genéticamente; Proteínas Recombinantes/biosíntesis; Proteínas Recombinantes/genética; Proteínas Recombinantes/aislamiento & purificación; Proteínas Recombinantes/farmacología; Factor de Células Madre/biosíntesis; Factor de Células Madre/genética; Factor de Células Madre/aislamiento & purificación; Nicotiana/metabolismo; Nicotiana/virología; Virus del Mosaico del Tabaco/genética; Transgenes

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Nicotiana / Factor I del Crecimiento Similar a la Insulina / Células Madre Hematopoyéticas / Diferenciación Celular / Eritropoyetina / Interleucina-3 / Factor de Células Madre Tipo de estudio: Guideline Idioma: En Revista: Stem Cells Dev Asunto de la revista: HEMATOLOGIA Año: 2013 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

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