Comparison of stem cell viability of bone marrow cryopreserved by two different methods |
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| Authors: | S Y Wang C K Ho P M Chen C H Yung L L Chong L Y Chen |
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| Institution: | 1. Department of Medical Research, Veterans General Hospital and National Yang-Ming Medical College, Taipei, Taiwan 112, Republic of China;2. Department of Internal Medicine, Veterans General Hospital and National Yang-Ming Medical College, Taipei, Taiwan 112, Republic of China;1. Department of Nutrition and Food Hygiene, Hubei Key Laboratory of Food Nutrition and Safety, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China;2. Ministry of Education Key Laboratory of Environment and Health, and State Key Laboratory of Environmental Health (Incubating), School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China;3. Department of Nutrition, Harvard TH Chan School of Public Health, Boston, MA, USA;4. Department of Epidemiology, Harvard TH Chan School of Public Health, Boston, MA, USA;5. CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China;6. Channing Division of Network Medicine, Department of Medicine, Brigham and Women''s Hospital and Harvard Medical School, Boston, MA, USA;7. Division of Preventive Medicine, Department of Medicine, Brigham and Women''s Hospital and Harvard Medical School, Boston, MA, USA;8. Division of Women''s Health, Department of Medicine, Brigham and Women''s Hospital and Harvard Medical School, Boston, MA, USA;1. Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey;2. Drug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia;3. Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan;4. Department of Field Crops, Faculty of Agriculture, Selcuk University, 42070 Konya, Turkey;1. Department of Orthopedic Surgery, VU University Medical Center, Amsterdam, The Netherlands;2. Department of Orthopedic Surgery, Noord-West Ziekenhuizen, Alkmaar, The Netherlands;3. MOVE Research Institute Amsterdam, Amsterdam, The Netherlands;4. Amsterdam Movement Sciences Institute, Amsterdam, The Netherlands;5. Faculty of Behavioral and Movement Sciences, VU University, Amsterdam, The Netherlands;6. Dept. of Medical Biology, Academisch Medisch Centrum, Amsterdam, The Netherlands;1. National Institute of Criminalistics and Criminology, Vilvoordsesteenweg 100, 1120 Brussels, Belgium;2. Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, 9000 Ghent, Belgium;3. Department of Laboratory Medicine, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium |
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| Abstract: | Two different cryogenic methods were used to study the preservation of murine bone marrow cells. Compared to the classical methods, in which separated mononuclear marrow cells in 10% dimethyl sulfoxide (DMSO) were cryopreserved in liquid nitrogen (-196 degrees C), a modified technique was carried out by cryopreservation of unfractionated marrow cells in a mixed protectant of 5% DMSO and 6% hydroxyethyl starch (HES) at -80 degrees C. Samples that were separately thawed after storage for 1, 4, 8, and 12 weeks were assayed for cell viability and recovery of CFU-GM and CFU-S. No macroscopic clumping of cells was noted either in fractionated or in unfractionated marrow cell cryopreservations. A mild damage, about 25% reduction of stem cells, was found at 1 week and did not deepen further. It seems that the greatest loss of stem cells occurred in the process of cryopreservation itself. Compared to prefreeze values, both a high number of cells that excluded trypan blue (87 +/- 3.4%) and a high recovery of CFU-GM (75 +/- 9.8%) and CFU-S (74 +/- 11.2) were observed in unfractionated marrow samples cryopreserved with the DMSO/HES mixture at -80 degrees C for 3 months and these results were very similar to those obtained from fractionated mononuclear marrow cells cryopreserved at -196 degrees C. The DMSO/HES protectant provides a simplified bone marrow cryopreservation technique that should be favorable to clinical application because of its high stem cell recovery and avoidance of cell-separation manipulation. |
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