| 骨髓基质干细胞-80℃保存的初步研究 |
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Received:November 30, 2006
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| 作者 | Author | 单位 | Unit | E-Mail |
| 蓝旭 |
LAN Xu |
解放军兰州军区总医院脊柱外科,甘肃兰州730050 |
Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China |
lzzyjw@sina.com |
| 文益民 |
WEN Yi-min |
解放军兰州军区总医院脊柱外科,甘肃兰州730050 |
Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China |
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| 葛宝丰 |
GE Bao-feng |
解放军兰州军区总医院脊柱外科,甘肃兰州730050 |
Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China |
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| 刘雪梅 |
LIU Xue-mei |
解放军兰州军区总医院脊柱外科,甘肃兰州730050 |
Department of Spine Surgery,the General Hospital of Lanzhou Military Command,Lanzhou 730050,Gansu,China |
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| 期刊信息：《中国骨伤》2007年20卷，第11期，第754-756页 |
| DOI：doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz |
| 基金项目:全军医学科学技术“十一五”计划课题基金资助项目(编号:06MA081) |
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目的:探索适合于人骨髓基质干细胞-80℃冷冻保存的条件和方法。
方法:体外分离培养人骨髓基质干细胞,以含不同浓度二甲亚砜的冻存液和不同细胞浓度-80℃深低温保存。3个月后复苏接种培养,观察细胞形态、存活率和贴壁率;通过3H-胸腺嘧啶脱氧核苷(3H-TdR)掺入实验比较细胞分裂增殖能力;3H-脯氨酸(3H-Proline)掺入实验检测胶原合成能力;RT-PCR法检测Ⅰ型胶原和骨钙素mRNA表达。
结果:不同二甲亚砜浓度保存骨髓基质干细胞复苏后的细胞形态和功能有所不同,其中含5%二甲亚砜冻存液组保存效果最差,15%组最佳。15%二甲亚砜冻存液组复苏后骨髓基质干细胞的形态与未冻存组相似,存活率为(85%±3%),贴壁率为(81%±5%)。骨髓基质干细胞保持较强的分裂增殖和胶原合成能力,Ⅰ型胶原和骨钙素mRNA表达与5%或10%二甲亚砜冻存液组比较有统计学差异(P<0.05)。冻存骨髓基质干细胞浓度为(5～10)×106个/ml组复苏后细胞存活率明显优于(1～2.5)×106个/ml组。
结论:含15%二甲亚砜的冻存液适合于骨髓基质干细胞长期保存,高浓度组骨髓基质干细胞冻存效果优于低浓度组。 |
| [关键词]:骨髓基质干细胞 细胞 培养的 低温保存 二甲亚砜 |
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| A preliminary study of cryopreservation of marrow stromal cells at-80℃ |
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Abstract:
Objective:To explore the condition and method for cryopreservation of marrow stromal cells(MSCs) at-80 ℃.
Methods:Human MSCs were isolated,cultured and stored in cryopreservation medium of different dimethylsulfoxide(DMSO) concentration and different concentration of MSCs at-80 ℃ in the condition of profound hypothermia.After 3 months cryopreservation,the MSCs were thawed and inoculated for culture.The morphology,viability and attachment rate of MSCs were observed.The proliferative ability of MSCs was detected by 3H-TdR incorporation test,the collagen synthesis was studied by 3H-Proline incorporation test,the mRNA expression of type Ⅰcollagen and osteocalcin was detected by RT-PCR technique.
Results:There were differences in cell morphology and function between different DMSO concentration groups for cryopreservation of MSCs.The preservation effect was the worst after stored in 5% DMSO group and best in 15% DMSO group.The morphology of MSCs stored in 15% DMSO group was similar with the group without cryopreservation.The cell viability rate was(85%±3%) and attachment rate was(81%±5%).MSCs kept the better proliferative ability and collagen synthesis.The difference of mRNA expression of type Ⅰcollagen and osteocalcin was significantly as compared with 5% and 10% DMSO group.The viability rate of MSCs in(5-10)×106/ml cryopreservation concentration was better than MSCs in(1-2.5)×106/ml.
Conclusion:15% DMSO cryopreservation solution is ideal for long-term preservation of MSCs,the cryopreservation effect of MSCs in high concentration is better than that in low concentration. |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 蓝旭,文益民,葛宝丰,刘雪梅.骨髓基质干细胞-80℃保存的初步研究[J].中国骨伤,2007,20(11):754~756 |
| 英文格式: | LAN Xu,WEN Yi-min,GE Bao-feng,LIU Xue-mei.A preliminary study of cryopreservation of marrow stromal cells at-80℃[J].zhongguo gu shang / China J Orthop Trauma ,2007,20(11):754~756 |
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