title: Selection of optimal passage of bone marrow-derived mesenchymal stem cells for stem cell therapy in patients with amyotrophic lateral sclerosis.
abstract: Mesenchymal stem cells (MSCs) obtained from bone marrow (BM) are currently used as an alternative therapy in amyotrophic lateral sclerosis (ALS) patients. Selection of optimal passages of autologous BM-derived MSCs during long-term in vitro expansion is important for clinical trials in patients with ALS. We isolated and expanded MSCs from the BM of eight ALS patients to analyze the growth kinetics, differentiation potential, cellular surface antigen expression, karyotype modifications and secretion of various cytokines during long-term culture. The morphology and size of the cells changed from small and spindle-like cells to large and polygonal types in later passages. The growth rate of the MSCs was highest in the third passage, followed by a gradual decrease. There were no special modifications of cell surface antigens or the karyotype of the MSCs from the first to the tenth passage. MSCs in the fourth passage were differentiated into adipocytes, osteocytes and chondrocytes. When we analyzed the cultured media of MSCs at the third, fifth, seventh and ninth passages, IL-6, VEGF and IL-8 showed high expression, with more than 50pg/10,000 cells at these passages; however, their expression progressively decreased with additional passages. In addition, secretion of IL-15, GM-CSF, IL-10, PDGF-bb, G-CSF, IL-1beta, basic FGF and IFN-gamma gradually decreased over prolonged culture. We suggest that MSCs at earlier passages are more suitable for stem cell therapy in ALS patients because of their stability and more potent anti-inflammatory and neuroprotective properties.
authors.affiliation: Division of Molecular and Life Sciences, Hanyang University, Ansan, 426-791, Republic of Korea.
authors.forename: Mi Ran
authors.initials: MR
authors.lastname: Choi
authors.forename: Hyun Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Ji-Yoon
authors.initials: JY
authors.lastname: Park
authors.forename: Tae Yong
authors.initials: TY
authors.lastname: Lee
authors.forename: Cheong Soon
authors.initials: CS
authors.lastname: Baik
authors.forename: Young Gyu
authors.initials: YG
authors.lastname: Chai
authors.forename: Kyoung Hwa
authors.initials: KH
authors.lastname: Jung
authors.forename: Kyoung Sun
authors.initials: KS
authors.lastname: Park
authors.forename: Wonjae
authors.initials: W
authors.lastname: Roh
authors.forename: Kyung Suk
authors.initials: KS
authors.lastname: Kim
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000628:therapy*; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D013997:Time Factors