title: Bone Marrow Mesenchymal Stromal Cells from Clinical Scale Culture: In Vitro Evaluation of Their Differentiation, Hematopoietic Support, and Immunosuppressive Capacities.
abstract: The differentiation capacity, hematopoietic support, and immunomodulatory properties of human bone marrow mesenchymal stromal cells (BM-MSCs) make them attractive therapeutic agents for a wide range of diseases. Clinical scale cultures (CSCs) have been used to expand BM-MSCs for their use in cell therapy protocols; however, little is known about the functionality of the expanded cells. The main goal of the present study was to evaluate the functional characteristics of BM-MSCs expanded from CSCs to determine the quality of the cells for cellular therapy protocols. To address this issue, we analyzed the morphology, immunophenotype, differentiation potential (adipogenic, osteogenic and chondrogenic), hematopoietic support, and immunosuppressive capacity of BM-MSCs from short scale cultures (SSCs) and CSCs in a comparative manner. After 12 days of culture in CSCs (HYPERFlask System), BM-MSCs reached cell numbers of 125.52 × 10(6) ± 25.6 × 10(6) MSCs, which corresponded to the number of cells required for transplantation (∼1.7 × 10(6) MSCs/kg for a 70-kg patient). After expansion, BM-MSCs expressed the characteristic markers CD73, CD90, and CD105; however, expansion decreased their differentiation capacity toward the adipogenic, osteogenic, and chondrogenic lineages and their ability to inhibit T-cell proliferation compared with SSCs-MSCs. Importantly, CSCs-MSCs maintained the ability to support the proliferation and expansion of hematopoietic progenitor cells and the capacity to express the molecules, cytokines, and extracellular matrix proteins involved in the regulation of hematopoiesis. Our study highlights the need to evaluate the functional properties of the expanded BM-MSCs for verification of their quality for cell therapy protocols.
authors.affiliation: 1 Mesenchymal Stem Cells Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .|2 Program in Biological Sciences of the National Autonomous University of Mexico (UNAM) , Mexico City, Mexico .
authors.forename: Guadalupe R
authors.initials: GR
authors.lastname: Fajardo-Orduña
authors.affiliation: 3 Hematopoietic Stem Cells Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Héctor
authors.initials: H
authors.lastname: Mayani
authors.affiliation: 4 CONACYT, Children Hospital of Mexico , Mexico City, Mexico .
authors.forename: Marta E
authors.initials: ME
authors.lastname: Castro-Manrreza
authors.affiliation: 5 Niche and Hematopoietic Microenvironment Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Eugenia
authors.initials: E
authors.lastname: Flores-Figueroa
authors.affiliation: 3 Hematopoietic Stem Cells Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Patricia
authors.initials: P
authors.lastname: Flores-Guzmán
authors.affiliation: 6 Immunochemistry Research Unit, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Lourdes
authors.initials: L
authors.lastname: Arriaga-Pizano
authors.affiliation: 7 Molecular Oncology Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Patricia
authors.initials: P
authors.lastname: Piña-Sánchez
authors.affiliation: 1 Mesenchymal Stem Cells Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Erika
authors.initials: E
authors.lastname: Hernández-Estévez
authors.affiliation: 8 Immunotherapy and Tissue Engineering Laboratory, Faculty of Medicine, UNAM , Mexico City, Mexico .
authors.forename: Andrés E
authors.initials: AE
authors.lastname: Castell-Rodríguez
authors.affiliation: 9 Immunology Research Unit, Pediatric Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Adriana K
authors.initials: AK
authors.lastname: Chávez-Rueda
authors.affiliation: 9 Immunology Research Unit, Pediatric Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: María V
authors.initials: MV
authors.lastname: Legorreta-Haquet
authors.affiliation: 10 Hematopoiesis and Leukemia Laboratory, FES-Zaragoza, UNAM , Mexico City, Mexico .
authors.forename: Edelmiro
authors.initials: E
authors.lastname: Santiago-Osorio
authors.affiliation: 1 Mesenchymal Stem Cells Laboratory, Oncology Research Unit, Oncology Hospital, National Medical Center , IMSS, Mexico City, Mexico .
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Montesinos
mesh_terms: D050156:Adipogenesis / Q000235:genetics; D015703:Antigens, CD / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D048430:Cell Shape / Q000235:genetics; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000235:genetics; D016207:Cytokines / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005786:Gene Expression Regulation; D006410:Hematopoiesis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007165:Immunosuppression Therapy; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D016207:Cytokines