A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement

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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...

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Autores principales:	Luzzani, Carlos Daniel, Questa, María, Guberman, Alejandra Sonia, Scassa, María Elida, Romorini, Leonardo
Publicado:	2015
Materias:	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 cytology drug effects fluorescence microscopy human embryonic stem cell mesenchymal stroma cell metabolism phenotype 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
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_17576512_v6_n1_p_Luzzani http://hdl.handle.net/20.500.12110/paper_17576512_v6_n1_p_Luzzani
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_17576512_v6_n1_p_Luzzani2023-06-08T16:28:57Z A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement Luzzani, Carlos Daniel Questa, María Guberman, Alejandra Sonia Scassa, María Elida Romorini, Leonardo 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 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. Fil:Luzzani, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Questa, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Guberman, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Scassa, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Romorini, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2015 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_17576512_v6_n1_p_Luzzani http://hdl.handle.net/20.500.12110/paper_17576512_v6_n1_p_Luzzani
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 Luzzani, Carlos Daniel Questa, María Guberman, Alejandra Sonia Scassa, María Elida Romorini, Leonardo A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
topic_facet	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
description	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.
author	Luzzani, Carlos Daniel Questa, María Guberman, Alejandra Sonia Scassa, María Elida Romorini, Leonardo
author_facet	Luzzani, Carlos Daniel Questa, María Guberman, Alejandra Sonia Scassa, María Elida Romorini, Leonardo
author_sort	Luzzani, Carlos Daniel
title	A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
title_short	A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
title_full	A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
title_fullStr	A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
title_full_unstemmed	A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
title_sort	therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement
publishDate	2015
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_17576512_v6_n1_p_Luzzani http://hdl.handle.net/20.500.12110/paper_17576512_v6_n1_p_Luzzani
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_version_	1768546463965511680

A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement

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