title: Long term expansion of bone marrow-derived hMSCs on novel synthetic microcarriers in xeno-free, defined conditions.
abstract: Human mesenchymal stem cells (hMSCs) present an attractive target for cell therapy given their wide availability, immunomodulatory properties, and multipotent nature for differentiation into chondrocytes, osteocytes, and adipocytes. With the progression of hMSC clinical studies, there is an increasing demand for development of technologies that enable efficient cell scale-up into clinically relevant quantities. Commercial scale manufacturing of hMSCs will require a large surface area which is not cost effective with available two-dimensional culture vessels. Recent studies showed that microcarriers provide a three-dimensional culture environment suitable for hMSC expansion. Traditionally, biological coatings and/or serum-containing medium are required to facilitate hMSC attachment and expansion in dynamic conditions. These limitations may hinder the use of microcarriers as a scale-up technology for hMSC therapeutics, where cell products, and therefore patient safety, are more controlled with the use of xeno-free, defined culture conditions. Here we report the long term culture of hMSCs on novel synthetic Synthemax II microcarriers in two different xeno-free media. Cells were maintained over 40 days on sterile, ready-to-use microcarriers in spinner flasks with programmed agitation. hMSC expansion was obtained by addition of fresh beads without the need for enzymatic dissociation. We achieved a cumulative cell expansion of >10,000 fold, and cells retained normal hMSC phenotype, karyotype, and tri-lineage differentiation potential. To our knowledge, this report is the first example of long term culture of hMSCs on synthetic microcarriers in xeno-free, defined conditions.
authors.affiliation: Corning European Technology Center, Corning Incorporated, Avon, France.
authors.forename: Martial
authors.initials: M
authors.lastname: Hervy
authors.affiliation: Corning Life Sciences Development, Corning Incorporated, Corning, New York, United States of America.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Weber
authors.affiliation: Corning European Technology Center, Corning Incorporated, Avon, France.
authors.forename: Marylene
authors.initials: M
authors.lastname: Pecheul
authors.affiliation: Corning Life Sciences Development, Corning Incorporated, Corning, New York, United States of America.
authors.forename: Paula
authors.initials: P
authors.lastname: Dolley-Sonneville
authors.affiliation: Corning European Technology Center, Corning Incorporated, Avon, France.
authors.forename: David
authors.initials: D
authors.lastname: Henry
authors.affiliation: Corning Life Sciences Development, Corning Incorporated, Corning, New York, United States of America.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Corning Life Sciences Development, Corning Incorporated, Corning, New York, United States of America.
authors.forename: Zara
authors.initials: Z
authors.lastname: Melkoumian
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology*; D006801:Humans; D059785:Karyotype; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008863:Microspheres; D010641:Phenotype; D013997:Time Factors
chemical_list: D015415:Biomarkers; D003470:Culture Media