title: Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo.
abstract: Mesenchymal stem cells (MSCs) possess great therapeutic potential. Efficient in vitro expansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrate for cell culture. In this study, we have aimed to improve the functionality and stemness of MSCs during culture using poly-L-lysine (PLL). Functionality of MSCs was analysed by cell cycle analysis, differentiation assay, β-galactosidase staining, and RT-PCR. Furthermore, we assessed the global gene expression profile of MSCs on uncoated and PLL-coated plates. MSCs on PLL-coated plates exhibited a faster growth rate with increased S-phase and upregulated expression of the stemness markers. In addition, their osteogenic differentiation potential was increased, and genes involved in cell adhesion, FGF-2 signalling, cell cycle, stemness, cell differentiation, and cell proliferation were upregulated, compared to that of the MSCs cultured on uncoated plates. We also confirmed that MSCs on uncoated plates expressed higher β-galactosidase than the MSCs on PLL-coated plates. We demonstrate that PLL provides favourable microenvironment for MSC culture by reversing the replicative senescence. This method will significantly contribute to effective preparation of MSCs for cellular therapy.
authors.affiliation: Cell Therapy Centre, Severance Hospital, Seoul 03722, Republic of Korea.
authors.forename: June Seok
authors.initials: JS
authors.lastname: Heo
authors.affiliation: Cell Therapy Centre, Severance Hospital, Seoul 03722, Republic of Korea; Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Hyun Ok
authors.initials: HO
authors.lastname: Kim
authors.affiliation: Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Seung Yong
authors.initials: SY
authors.lastname: Song
authors.affiliation: Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Dae Hyun
authors.initials: DH
authors.lastname: Lew
authors.affiliation: Cell Therapy Centre, Severance Hospital, Seoul 03722, Republic of Korea.
authors.forename: Youjeong
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.
authors.forename: Sinyoung
authors.identifier: 0000-0002-2609-8945
authors.initials: S
authors.lastname: Kim
mesh_terms: D017667:Adipocytes / Q000166:cytology; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D005109:Extracellular Matrix / Q000378:metabolism; D005434:Flow Cytometry; D015894:Genome, Human; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D020411:Oligonucleotide Array Sequence Analysis; D010012:Osteogenesis; D011107:Polylysine / Q000737:chemistry*; D044968:Regenerative Medicine; D016196:S Phase; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D011107:Polylysine; D001616:beta-Galactosidase