Selection of Optimal M esenchymal Stem Cells for Stem Cell Therapy in Amyotrophic Lateral Sclerosis Patients

Abstract

Background & Objectives: Mesenchymal stem cells (MSC) from bone marrow have been used as an alternative therapy for amyotrophic lateral sclerosis (ALS) patients. However, the stability and characterization of MSCs during long-term in vitro expansion, which is essential for clinical trials, have not yet been evaluated.

Method: We isolated and expanded MSCs from bone marrow of eight ALS patients to analyze the growth kinetics, cellular surface antigen, differentiation potential, karyotype modifications, and the secretion of various cytokines and growth factors during long-term culture of MSCs.

Results: The morphology and size of the cells transitioned from small and spindle-like cells to large and polygonal types in later passages. The growth rate of the MSCs was highest in the 3rd passage (3.15±0.65) and then gradually decreased. There were no special modifications of cell surface antigens (CD29, CD44, CD73, CD105, CD34, CD45, and HLA-DR) or the karyotype of the MSCs from the 1st to 10th passage. MSCs in the 4th passage were induced to differentiate into mesenchymal lineage cells, including adipocytes, osteocytes, and chondrocytes. When we analyzed the cultured media of MSCs at 3rd, 5th, 7th, and 9th passages, IL-6, VEGF, and IL-8 showed high expression, with more than 50 pg/10,000 cells in all these passages, but their expression continuously decreased with time. Additionally, secretion of IL-15, GM-CSF, IL-10, PDGF bb, G-CSF, IL-1β, FGF basic, and IFN-gamma gradually decreased as culture time progressed.

Conclusion: We therefore suggest that MSCs in earlier passages are suitable for stem cell therapy because of their stability and neuroprotective roles associated with various cytokines and growth factors.