多发性骨髓瘤患者骨髓间充质干细胞冻存前后的生物学特性

摘要

背景：多发性骨髓瘤患者的骨髓间充质干细胞具有多向分化、免疫调节和支持造血作用，但是这些功能是否受冻存的影响目前尚不清楚。　　目的：探讨冻存对多发性骨髓瘤患者骨髓间充质干细胞生物学特性的影响。　　方法：采用细胞贴壁法获取多发性骨髓瘤患者骨髓间充质干细胞，将传代后的细胞用 IMDM 细胞冻存液(含10%的二甲基亚砜和体积分数40%的胎牛血清)保存在-196℃液氮中。检测短期(1个月)和长期(12个月)冻存复苏后间充质干细胞的活性和增殖能力；将冻存后多发性骨髓瘤患者骨髓间充质干细胞作为滋养层，应用甲基纤维素半固体培养，检测其支持造血的能力；混合淋巴细胞反应检测冻存后多发性骨髓瘤患者骨髓间充质干细胞调控免疫能力。　　结果与结论：经过短、长期冻存后多发性骨髓瘤患者骨髓间充质干细胞的细胞活性分别为(92.9±7.5)%和(86.7±9.2)%；短、长期冻存后细胞的增殖能力与冻存前间充质干细胞相似；冻存后多发性骨髓瘤患者骨髓间充质干细胞仍具有支持造血祖细胞生长的作用和抑制T淋巴细胞增殖的能力，与冻存前相比，没有明显差别。说明冻存可以降低多发性骨髓瘤患者骨髓间充质干细胞的细胞活性，但是并不影响间充质干细胞的增殖、支持造血和免疫调节的能力。%BACKGROUND:Bone marrow mesenchymal stem cells derived from multiple myeloma patients are characterized by pluripotential differentiation, immunoregulation and supporting hematopoiesis, but whether these features are affected after cryopreservation is unclear. OBJECTIVE:To study the biological characteristics of cryopreserved bone marrow mesenthymal stem cellderived from multiple myeloma patients. METHODS:Bone marrow mesenchymal stem cells derived from multiple myeloma patients were cryopreserved in-196 ℃ liquid nitrogen for 1 month (short term) and 12 months (long term) with Iscove’s modified Dulbecco’s medium containing 10%dimethyl sulfoxide and 40%fetal bovine serum as cryoprotectant. The viability and proliferation ability of thawed bone marrow mesenchymal stem cells were investigated. Hematopoiesis support of thawed bone marrow mesenchymal stem cells in vitro was detected by long-term bone marrow culture and the methylcellulose progenitor assay. The immunoregulatory ability of thawed mesenchymal stem cells was detected by mixed lymphocyte culture assay. RESULTS AND CONCLUSION:The cellviability was (92.9±7.5)%and (86.7±9.2)%for mesenchymal stem cells cryopreserved as long as 1 month or 12 months, respectively. Furthermore, thawed bone marrow mesenchymal stem cells possessed the ability of supporting colony forming and could significantly suppress proliferation of T lymphocytes. At last, there were no changes detected as compared with pre-cryopreserved mesenchymal stem cells in the abilities of proliferation, hematopoiesis support and immunoregulation. Cryopreservation can decrease the cellviability of bone marrow mesenchymal stem cells derived from the patients with multiple myeloma, but cannot affect the abilities of proliferation, hematopoiesis support and immunoregulation.