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Luzzani, C.; Neiman, G.; Garate, X.; Questa, M.; Solari, C.; Fernandez Espinosa, D.; García, M.; Errecalde, A.L.; Guberman, A.; Scassa, M.E.; Sevlever, G.E.; Romorini, L.; Miriuka, S.G. "A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement" (2015) Stem Cell Research and Therapy. 6(1)
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Abstract:

Introduction: Mesenchymal stem cells (MSCs) are a promising source of cells for regenerative therapies. Although they can be isolated easily from several tissues, cell expansion is limited since their properties are lost with successive passages. Hence, pluripotent derived MSCs (PD-MSCs) arise as a suitable alternative for MSC production. Nevertheless, at present, PD-MSC derivation protocols are either expensive or not suitable for clinical purposes. Methods: In this work we present a therapy-grade, inexpensive and simple protocol to derive MSCs from pluripotent stem cells (PSCs) based on the use of platelet lysate (PL) as medium supplement. Results: We showed that the PD-MSCPL expressed multiple MSC markers, including CD90, CD73, CD105, CD166, and CD271, among others. These cells also show multilineage differentiation ability and immunomodulatory effects on pre-stimulated lymphocytes. Thorough characterization of these cells showed that a PD-MSCPL resembles an umbilical cord (UC) MSC and differs from a PSC in surface marker and extracellular matrix proteins and integrin expression. Moreover, the OCT-4 promoter is re-methylated with mesenchymal differentiation comparable with the methylation levels of UC-MSCs and fibroblasts. Lastly, the use of PL-supplemented medium generates significantly more MSCs than the use of fetal bovine serum. Conclusions: This protocol can be used to generate a large amount of PD-MSCs with low cost and is compatible with clinical therapies. © 2015 Luzzani et al.; licensee BioMed Central.

Registro:

Documento: Artículo
Título:A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
Autor:Luzzani, C.; Neiman, G.; Garate, X.; Questa, M.; Solari, C.; Fernandez Espinosa, D.; García, M.; Errecalde, A.L.; Guberman, A.; Scassa, M.E.; Sevlever, G.E.; Romorini, L.; Miriuka, S.G.
Filiación:Laboratorio de Biología Del Desarrollo Celular, LIAN-Unidad Asociada Al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina
Laboratorio de Regulación de Expresión Génica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Buenos Aires, Argentina
Cátedra de Citología, Histología y Embriología, Facultad de Ciencias Médicas, Universidad Nacional de la Plata, Calle 60 s/n, La Plata, 1900, Argentina
Investigador, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina
Palabras clave:5' nucleotidase; activated leukocyte cell adhesion molecule; CD271 antigen; cell surface marker; endoglin; integrin; lymphocyte antigen; octamer transcription factor 4; scleroprotein; Thy 1 antigen; unclassified drug; membrane antigen; signal peptide; antigen expression; Article; cell differentiation; cell lineage; cell lysate; cell stimulation; controlled study; culture medium; DNA methylation; embryo; flow cytometry; human; human cell; immunomodulation; lymphocyte; mesenchymal stem cell; peripheral blood mononuclear cell; platelet lysate; pluripotent stem cell; priority journal; promoter region; protein analysis; protein expression; stem cell expansion; umbilical cord; cell culture; cell differentiation; cytology; drug effects; fluorescence microscopy; human embryonic stem cell; mesenchymal stroma cell; metabolism; phenotype; pluripotent stem cell; thrombocyte; Bovinae; Antigens, Surface; Blood Platelets; Cell Differentiation; Cells, Cultured; DNA Methylation; Human Embryonic Stem Cells; Humans; Intercellular Signaling Peptides and Proteins; Mesenchymal Stromal Cells; Microscopy, Fluorescence; Phenotype; Pluripotent Stem Cells; Promoter Regions, Genetic
Año:2015
Volumen:6
Número:1
DOI: http://dx.doi.org/10.1186/scrt540
Título revista:Stem Cell Research and Therapy
Título revista abreviado:Stem Cell Res. Ther.
ISSN:17576512
CAS:5' nucleotidase, 9027-73-0; Antigens, Surface; Intercellular Signaling Peptides and Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_17576512_v6_n1_p_Luzzani

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Citas:

---------- APA ----------
Luzzani, C., Neiman, G., Garate, X., Questa, M., Solari, C., Fernandez Espinosa, D., García, M.,..., Miriuka, S.G. (2015) . A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement. Stem Cell Research and Therapy, 6(1).
http://dx.doi.org/10.1186/scrt540
---------- CHICAGO ----------
Luzzani, C., Neiman, G., Garate, X., Questa, M., Solari, C., Fernandez Espinosa, D., et al. "A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement" . Stem Cell Research and Therapy 6, no. 1 (2015).
http://dx.doi.org/10.1186/scrt540
---------- MLA ----------
Luzzani, C., Neiman, G., Garate, X., Questa, M., Solari, C., Fernandez Espinosa, D., et al. "A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement" . Stem Cell Research and Therapy, vol. 6, no. 1, 2015.
http://dx.doi.org/10.1186/scrt540
---------- VANCOUVER ----------
Luzzani, C., Neiman, G., Garate, X., Questa, M., Solari, C., Fernandez Espinosa, D., et al. A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement. Stem Cell Res. Ther. 2015;6(1).
http://dx.doi.org/10.1186/scrt540