title: Genomic and functional comparison of mesenchymal stromal cells prepared using two isolation methods.
abstract: OBJECTIVE
  Mesenchymal stromal cells (MSCs) have been applied to patients in cell therapy for various diseases. Recently, we introduced a novel MSC separation filter device which could yield approximately 2.5-fold more MSCs from bone marrow in a closed system compared with the conventional open density gradient centrifugation method. MSCs isolated with these two methods were phenotypically similar and met the criteria defining human MSC proposed by the International Society for Cellular Therapy. However, these criteria do not reflect the functional capacity of MSCs. It has been shown that the donor, source, isolation method, culture condition and cryopreservation of MSCs have potential to alter their therapeutic efficacy. To determine the equivalency of MSCs isolated by these two methods, we compared their genomic profiles as an index of their biologic potential and evaluated their growth promoting potential as an index of function.
  
  
  METHODS
  The gene expression profiles of human MSCs isolated from 5 healthy donors with two distinct methods were obtained from microarray analyses. The functional activity of freshly expanded/cryopreserved MSCs from these two isolation methods was evaluated using an in vitro chondrocyte proliferation assay.
  
  
  RESULTS
  Freshly expanded MSCs isolated by these two methods were found to exhibit similar gene expression profiles and equivalent therapeutic effects, while freshly thawed, cryopreserved MSCs lacked all measureable therapeutic activity.
  
  
  CONCLUSIONS
  The MSC separation device generates genomically and functionally equivalent MSCs compared with the conventionally isolated MSCs, although freshly thawed, cryopreserved MSCs, isolated by either method, are devoid of activity in our bioassay.
authors: Division of Oncology/Blood and Marrow Transplantation, The Children's Hospital of Philadelphia and The University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA; Center for Childhood Cancer and Blood Diseases, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA.
authors: Satoru
authors: S
authors: Otsuru
authors: Division of Oncology/Blood and Marrow Transplantation, The Children's Hospital of Philadelphia and The University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA.
authors: Ted J
authors: TJ
authors: Hofmann
authors: Center for Biomedical Informatics, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
authors: Pichai
authors: P
authors: Raman
authors: Division of Oncology/Blood and Marrow Transplantation, The Children's Hospital of Philadelphia and The University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA.
authors: Timothy S
authors: TS
authors: Olson
authors: Center for Childhood Cancer and Blood Diseases, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA.
authors: Adam J
authors: AJ
authors: Guess
authors: Department of Oncology and Hematology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Massimo
authors: M
authors: Dominici
authors: Division of Oncology/Blood and Marrow Transplantation, The Children's Hospital of Philadelphia and The University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA; Center for Childhood Cancer and Blood Diseases, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, USA; Divsion of Hematology/Oncology/BMT, Nationwide Children's Hospital, Columbus, Ohio, USA. Electronic address: edwin.horwitz@nationwidechildrens.org.
authors: Edwin M
authors: EM
authors: Horwitz
mesh_terms: D000328:Adult; D000818:Animals; D001853:Bone Marrow; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D020869:Gene Expression Profiling; D023281:Genomics; D006801:Humans; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D014183:Transplantation, Heterologous; D055815:Young Adult
keywords: bone marrow MSC; cryopreservation; functional assay; gene expression profile