인간 골수유래 중간엽 줄기세포의 대량생산을 위한 새로운 배양 시스템.

Abstract

Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation and have potential applications in a range of therapies. MSCs obtained from bone marrow (BM) are currently used as an alternative therapy in intractable neurological disorders. Extremely higher numbers of stem cells are needed for clinical trials (up to millions of cells/kg of body weight). Current conventional culture methods are labor intensive procedure and have disadvantage in cost benefit issues. The main goal of the present study was to develop a new culture system for the efficient expansion of hBM-MSCs. At first, we studied effects of various media volume (0.15, 0.3, 0.5 ml/cm2) in static culture condition to increase cell yield. The results showed that total cell number was increased 1.3 fold corresponding to media volume. Population doubling time (PDT) at 0.5 ml/cm2 of media volume was shortened for 21 hours compared another conditions. Average lactate production rate was 1.1 mM. We developed new culture system, Garnet Project-1 (GP-1) which is modified roller bottle culture system for the efficient expansion of hBM-MSCs aspects of cost-effectiveness. The results showed that total cell number was increased 2 fold. Population doubling time (PDT) at GP-1 was 58 hours compared with 74 hours at control. MSCs in GP-1 culture system expressed the characteristic markers CD29, CD44, CD73, CD49C and CD105, whereas was negative for human leukocyte antigen (HLA)-DR. Moreover, Lactate production in 10 days was significantly 1.7 fold decrease in GP-1 compared to conventional culture. These results demonstrated the feasibility of expanding human BM-MSCs in a GP-1 culture system and represent an important step forward for the implementation of a Good Manufacturing Practices-compliant large-scale production system of hBM-MSCs for cellular therapy.