title: Identification and validation of multiple cell surface markers of clinical-grade adipose-derived mesenchymal stromal cells as novel release criteria for good manufacturing practice-compliant production.
abstract: Clinical translation of mesenchymal stromal cells (MSCs) necessitates basic characterization of the cell product since variability in biological source and processing of MSCs may impact therapeutic outcomes. Although expression of classical cell surface markers (e.g., CD90, CD73, CD105, and CD44) is used to define MSCs, identification of functionally relevant cell surface markers would provide more robust release criteria and options for quality control. In addition, cell surface expression may distinguish between MSCs from different sources, including bone marrow-derived MSCs and clinical-grade adipose-derived MSCs (AMSCs) grown in human platelet lysate (hPL).
  
  
  
  In this work we utilized quantitative PCR, flow cytometry, and RNA-sequencing to characterize AMSCs grown in hPL and validated non-classical markers in 15 clinical-grade donors.
  
  
  
  We characterized the surface marker transcriptome of AMSCs, validated the expression of classical markers, and identified nine non-classical markers (i.e., CD36, CD163, CD271, CD200, CD273, CD274, CD146, CD248, and CD140B) that may potentially discriminate AMSCs from other cell types. More importantly, these markers exhibit variability in cell surface expression among different cell isolates from a diverse cohort of donors, including freshly prepared, previously frozen, or proliferative state AMSCs and may be informative when manufacturing cells.
  
  
  
  Our study establishes that clinical-grade AMSCs expanded in hPL represent a homogeneous cell culture population according to classical markers,. Additionally, we validated new biomarkers for further AMSC characterization that may provide novel information guiding the development of new release criteria.
  
  
  
  Use of Autologous Bone Marrow Aspirate Concentrate in Painful Knee Osteoarthritis (BMAC): Clinicaltrials.gov NCT01931007 . Registered August 26, 2013. MSC for Occlusive Disease of the Kidney: Clinicaltrials.gov NCT01840540 . Registered April 23, 2013. Mesenchymal Stem Cell Therapy in Multiple System Atrophy: Clinicaltrials.gov NCT02315027 . Registered October 31, 2014. Efficacy and Safety of Adult Human Mesenchymal Stem Cells to Treat Steroid Refractory Acute Graft Versus Host Disease. Clinicaltrials.gov NCT00366145 . Registered August 17, 2006. A Dose-escalation Safety Trial for Intrathecal Autologous Mesenchymal Stem Cell Therapy in Amyotrophic Lateral Sclerosis. Clinicaltrials.gov NCT01609283 . Registered May 18, 2012.
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Emily T
authors: ET
authors: Camilleri
authors: Department of Laboratory Medicine & Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Michael P
authors: MP
authors: Gustafson
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Amel
authors: A
authors: Dudakovic
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Scott M
authors: SM
authors: Riester
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Catalina Galeano
authors: CG
authors: Garces
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Christopher R
authors: CR
authors: Paradise
authors: Department of Periodontology, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
authors: Hideki
authors: H
authors: Takai
authors: Department of Developmental Bioengineering, University of Twente, Enschede, The Netherlands.|Department of Tissue Regeneration, University of Twente, Enschede, The Netherlands.
authors: Marcel
authors: M
authors: Karperien
authors: Institute of Medical Biology, Agency for Science, Technology and Research, Singapore, Singapore.|Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors: Simon
authors: S
authors: Cool
authors: Department of Biochemistry, Rush University Medical Center, Chicago, IL, USA.|Department of Orthopedic Surgery, Rush University Medical Center, Chicago, IL, USA.|Department of Internal Medicine, Section of Rheumatology, Rush University Medical Center, Chicago, IL, USA.|Jesse Brown VA Medical Center, Chicago, IL, USA.
authors: Hee-Jeong Im
authors: HJ
authors: Sampen
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: A Noelle
authors: AN
authors: Larson
authors: Department of Physical Medicine and Rehabilitation, Division of Pain Medicine, Mayo Clinic, Rochester, MN, USA.
authors: Wenchun
authors: W
authors: Qu
authors: Department of Physical Medicine and Rehabilitation, Mayo Clinic, Rochester, MN, USA.|Department of Radiology, Mayo Clinic, Rochester, MN, USA.|Department of Anatomy, Mayo Clinic, Rochester, MN, USA.
authors: Jay
authors: J
authors: Smith
authors: Department of Laboratory Medicine & Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Allan B
authors: AB
authors: Dietz
authors: Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA. vanwijnen.andre@mayo.edu.|Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA. vanwijnen.andre@mayo.edu.
authors: Andre J
authors: AJ
authors: van Wijnen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D050154:Adiposity / Q000502:physiology; D015415:Biomarkers / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D017423:Sequence Analysis, RNA / Q000379:methods; D059467:Transcriptome / Q000502:physiology
chemical_list: D015415:Biomarkers
keywords: Adipose-derived mesenchymal stromal cells; CD markers; Flow cytometry; Human platelet lysate; Manufacturing; RNA-sequencing; Release criteria