Chapter 19 - Cryopreservation of mesenchymal stem cells (MSCs) derived from bone marrow with carbohydrate additive sucrose and dimethyl sulfoxide (DMSO)

Abstract

Effective cryopreservation strategies for mesenchymal stem cells (MSCs) are highly essential to be used in future “off-the-shelf” therapeutic purposes. The bone marrow-derived MSCs (BM-MSCs) are still considered as safest source for therapeutic intervention. However, the convectional cryoprotectant (CPA) dimethyl sulfoxide (DMSO) is known to have toxic side effects. So, it is always desirable to develop preservation medium for stem cells without or with reduced concentration of DMSO. In this study, we reduced the concentration of DMSO by addition of nontoxic CPA sucrose. Here, the combination of sucrose (0.015 M) and DMSO (7.5%) demonstrated better post-thaw survival rate (86.078 ± 0.001)% than DMSO at concentration 10% (81.238 ± 0.001)% and 7.5% (78.908 ± 0.001) for BM-MSCs after stored for 60 days. The population doubling time (PDT) of BM-MSCs cryopreserved along with 7.5% DMSO plus 0.015 M sucrose was found (33.244 ± 10.526) h similar to noncryopreserved BM-MSCs (30.08 ± 10.22) h. However, longer PDT was recorded when BM-MSCs were cryopreservation done in 10% DMSO (38.241 ± 11.908) h and 7.5% DMSO (43.102 ± 14.821) h. Our results indicate the post-thawed MSCs were able to generate MSC-like colonies, retain osteogenic and adipogenic differentiation capacity. In this report we develop a user friendly cryopreservation medium for long-term storage of BM-MSCs using sucrose that chiefly responsible to reduce the concentration of DMSO. So, that negative effect of DMSO can be minimized and post-transplantation complication may be evaded.