title: Enhanced ex vivo expansion of adult mesenchymal stem cells by fetal mesenchymal stem cell ECM.
abstract: Large-scale expansion of highly functional adult human mesenchymal stem cells (aMSCs) remains technologically challenging as aMSCs lose self renewal capacity and multipotency during traditional long-term culture and their quality/quantity declines with donor age and disease. Identification of culture conditions enabling prolonged expansion and rejuvenation would have dramatic impact in regenerative medicine. aMSC-derived decellularized extracellular matrix (ECM) has been shown to provide such microenvironment which promotes MSC self renewal and "stemness". Since previous studies have demonstrated superior proliferation and osteogenic potential of human fetal MSCs (fMSCs), we hypothesize that their ECM may promote expansion of clinically relevant aMSCs. We demonstrated that aMSCs were more proliferative (∼ 1.6 ×) on fMSC-derived ECM than aMSC-derived ECMs and traditional tissue culture wares (TCPS). These aMSCs were smaller and more uniform in size (median ± interquartile range: 15.5 ± 4.1 μm versus 17.2 ± 5.0 μm and 15.5 ± 4.1 μm for aMSC ECM and TCPS respectively), exhibited the necessary biomarker signatures, and stained positive for osteogenic, adipogenic and chondrogenic expressions; indications that they maintained multipotency during culture. Furthermore, fMSC ECM improved the proliferation (∼ 2.2 ×), size (19.6 ± 11.9 μm vs 30.2 ± 14.5 μm) and differentiation potential in late-passaged aMSCs compared to TCPS. In conclusion, we have established fMSC ECM as a promising cell culture platform for ex vivo expansion of aMSCs.
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore. Electronic address: ng.chee.ping@smart.mit.edu.
authors.forename: Chee Ping
authors.initials: CP
authors.lastname: Ng
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore.
authors.forename: Abdul Rahim Mohamed
authors.initials: AR
authors.lastname: Sharif
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Heath
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore.
authors.forename: John W
authors.initials: JW
authors.lastname: Chow
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore.
authors.forename: Claire B Y
authors.initials: CB
authors.lastname: Zhang
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459, Singapore.
authors.forename: Mary B
authors.initials: MB
authors.lastname: Chan-Park
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore; Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Paula T
authors.initials: PT
authors.lastname: Hammond
authors.affiliation: Experimental Fetal Medicine Group, Department of Obstetrics & Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore and National University Hospital Systems, Singapore, 5 Lower Kent Ridge Rd, Singapore 119074, Singapore; Department of Reproductive Medicine, KK Women's and Children's Hospital, 100 Bukit Timah Road, Singapore 229899, Singapore; Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, 8 College Road, Singapore 169857, Singapore.
authors.forename: Jerry K Y
authors.initials: JK
authors.lastname: Chan
authors.affiliation: BioSystems and Micromechanics Interdisciplinary Group, Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, Singapore 138602, Singapore; Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; Department of Mechanical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Linda G
authors.initials: LG
authors.lastname: Griffith
mesh_terms: D040761:Biomimetic Materials; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*
keywords: Bioactivity; Biomimetic material; Bone marrow; Cell proliferation; ECM (extracellular matrix); Mesenchymal stem cells