title: A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement.
abstract: BACKGROUND
  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.
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. cluzzani@fleni.org.ar.
authors.forename: Carlos
authors.initials: C
authors.lastname: Luzzani
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. gabrielneiman@hotmail.com.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Neiman
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. ximena.garate@gmail.com.
authors.forename: Ximena
authors.initials: X
authors.lastname: Garate
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. mariaquesta@gmail.com.
authors.forename: María
authors.initials: M
authors.lastname: Questa
authors.affiliation: Laboratorio de Regulación de Expresión Génica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Buenos Aires, Argentina. cmsolari@qb.fcen.uba.ar.
authors.forename: Claudia
authors.initials: C
authors.lastname: Solari
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. dfernandez@fleni.org.ar.
authors.forename: Darío
authors.initials: D
authors.lastname: Fernandez Espinosa
authors.affiliation: Cátedra de Citología, Histología y Embriología, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Calle 60 s/n, 1900, La Plata, Argentina. mngarcia@med.unlp.edu.ar.
authors.forename: Marcela
authors.initials: M
authors.lastname: García
authors.affiliation: Cátedra de Citología, Histología y Embriología, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Calle 60 s/n, 1900, La Plata, Argentina. aerreca@med.unlp.edu.ar.
authors.forename: Ana Lía
authors.initials: AL
authors.lastname: Errecalde
authors.affiliation: Laboratorio de Regulación de Expresión Génica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Intendente Güiraldes 2160, Buenos Aires, Argentina. algub@qb.fcen.uba.ar.|Investigador, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. algub@qb.fcen.uba.ar.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Guberman
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. mescassa@fleni.org.ar.
authors.forename: María Elida
authors.initials: ME
authors.lastname: Scassa
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. gsevlever@gmail.com.
authors.forename: Gustavo Emilio
authors.initials: GE
authors.lastname: Sevlever
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. lromorini@fleni.org.ar.|Investigador, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. lromorini@fleni.org.ar.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Romorini
authors.affiliation: Laboratorio de Biología del Desarrollo Celular, LIAN-Unidad Asociada al CONICET, Fundación FLENI, Ruta 9, Km53, Belen de Escobar, Argentina. smiriuka@fleni.org.ar.|Cátedra de Citología, Histología y Embriología, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Calle 60 s/n, 1900, La Plata, Argentina. smiriuka@fleni.org.ar.|Investigador, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. smiriuka@fleni.org.ar.
authors.forename: Santiago Gabriel
authors.initials: SG
authors.lastname: Miriuka
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019175:DNA Methylation; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic
chemical_list: D000954:Antigens, Surface; D036341:Intercellular Signaling Peptides and Proteins