| The Improvement of Cell Viability due to Dilution and Removal of DMSO in Thawing of Stem Cells |
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Hoon-Kyung Lee1, Kyung-Ha Ryu1, Il-Tae Whang1, Eun Suk Kang2, Ki Sook Hong2, Kyung-Hyo Kim1, Young-Mi Hong1, Kyung-Hee Kim1, Keun Lee1 |
1Departments of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Korea
2Departments of Clinical Pathology, College of Medicine, Ewha Womans University, Seoul, Korea |
| 조혈세포 해동 시 삼투압 희석과 DMSO 제거 방법을 이용한 세포 생존율 향상에 관한 연구 |
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이훈경1, 유경하1, 황일태1, 강은숙2, 홍기숙2, 김경효1, 홍영미1, 김경희1, 이근1 |
1이화여자대학교 의과대학 소아과학교실
2이화여자대학교 의과대학 임상병리학과교실 |
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| Abstract |
Purpose
: Cryopreserved stem cells from cord blood are usually infused with Dimethyl Sulfoxide (DMSO) immediately after thawing. However, this process may cause cell damage due to osmotic shock, and the administration of DMSO may also cause toxic effects. We studied a new method of increasing cell viability by stabilizing osmolarity by adding dextran 40 and washing out DMSO.
Methods
: Thirty-five samples of cord blood were studied. RBCs were removed in 10% pentastarch. The cells were mixed with DMSO of 5, 10 and 20% each, and stored at -80℃. Cryopreserved cells were thawed and then diluted with dextran 40. DMSO was removed afterwards. The cell viability, osmolarity and colony forming capacity in this new thawing method were compared with the control group done by conventional methods.
Results
: The recovery rate of WBC after RBCs separation was 92.06% but the contamination rate of RBC was still high(29.90%). The concentration of DMSO significantly affected the survival of WBCs(P<0.05). The osmolarity was 330±17.7mOsm/L before freezing, 1,457±508.7mOsm/L after thawing prior to dilution and 811±199.6mOsm/L after dilution, suggesting that the dilution process was effective in reducing osmolarity. The number of viable cells increased from 6.01±1.61(×103/L) to 7.16±1.48(×103/L) after dilution but was not significant. The number of CFU-C was increased from 5.82±4.19(/105) to 7.58±3.16(/105) after dilution but was not significant.
Conclusion
: Our method of removing DMSO during the thawing process yields a higher cell survival rate and less DMSO toxicities compared with the conventional method of direct injection with DMSO after thawing. |
| Key Words:
Stem cell, Dilution process, Viable cells, DMSO |
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