不同冻存方法对骨髓基质细胞生物学活性的影响  

Effects of Different Cryopreservation Methods on the Biological Properties of Bone Marrow Stromal Cells(BMSc)

作　　者：李厚轩[1] 闫福华[1] 刘建国[2] 林敏魁[1] 詹暶[1] 赵欣[1] 

机构地区：[1]福建医科大学口腔医学院牙周科,福建福州350002 [2]遵义医学院口腔医学系

出　　处：《口腔医学研究》2005年第6期604-608,共5页Journal of Oral Science Research

基　　金：国家自然科学基金资助项目(编号:30471892);福建省科技厅重大课题资助项目(编号:2001Z021)

摘　　要：目的:比较不同冻存方法对骨髓基质细胞(BMSc)增殖分化能力的影响。方法:从Beagle犬股骨抽取骨髓,培养,传代,至第4代时将其以不同细胞浓度(1×107/L、1×108/L、1×109/L)、不同浓度冻存保护剂DMSO(5%、10%、15%)、不同降温方式以及消化或原位等方法进行冻存,细胞复苏后传至第7代,观察冻存对其增殖分化能力的影响。结果:各种方法冻存的BMSc均保持了较高的增殖、分化能力。高细胞浓度、10%DMSO的冻存保护剂有利于细胞活性的保存(P<0.05),3种降温方式对BMSc的生物特性的影响没有显著差别。-80℃原位冻存时BMSc仍保存了较高的增殖、分化能力。结论:较高的冻存细胞浓度、选用含10%DMSO的冻存保护剂、适宜的降温方式有利于冻存。-80℃原位冻存可作为BMSc短期的保存方法。Objective： To observe the effects of various cryopreservation methods on the biological characteristics of bone marrow stromal cells （BMSc）. Methods： BMSc was harvested from the femur of beagle dog, then cultured and passaged. The fourth passage BMSc were cryopreserved at -80℃ in situ - 196℃ （ liquid nitrogen） in digested groups with different cell concentrations（ 1 × 10^7/L,1 × 10^8/L,1 × 10^9/L）, cooling rates, and culture medium （DMEM） containing 5%, 10% or 15% DMSO. They were recovered after one month to investigate whether the cryopreservation changed their potential of proliferation and differentiation. Results： The proliferation and differentiation potential of BMSc was rather good after recovery of cryopreservation. High cell concentrations and 10% DMSO were better choices for cryopreservation （P 〈 0.05 ）. The three cooling rates had no significant different effect on BMSc. Conclusion： High cell concentration, culture medium （DMEM） containing 10% DMSO might be favorite means for the cryopreservation of the BMSc. BMSc might be persevered at -80℃ in situ in one month.

关 键 词：骨髓基质细胞 冻存 复苏 增殖 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]