Biological characteristics of bone marrow mesenchymal stem cells from multiple myeloma patients before and after cryopreservation

doi:10.3969/j.issn.2095-4344.2013.45.012

Abstract

Abstract:

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 cell derived 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 cell viability 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 cell viability of bone marrow mesenchymal stem cells derived from the patients with multiple myeloma, but cannot affect the abilities of proliferation, hematopoiesis support and immunoregulation.

Key words: stem cells, multiple myeloma, mesenchymal stem cells, cryopreservation, freezing

CLC Number:

R394.2

Wang Zhen-ling, Wang Jin-huan, Guo Qing, Zhao Zhi-gang. Biological characteristics of bone marrow mesenchymal stem cells from multiple myeloma patients before and after cryopreservation[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2013.45.012.

Figures/Tables 3

2.1 冻存后骨髓间充质干细胞的形态和生长特点冻存前后多发性骨髓瘤患者骨髓具有相似的形态。倒置显微镜下多发性骨髓瘤患者骨髓间充质干细胞胞浆丰富，核染色质细，核仁明显，平行排列或“漩涡”样生长。根据细胞生长曲线，间充质干细胞的倍增时间为(47.6±5.7) h。细胞形态和倍增时间随着细胞传代并不发生改变。 2.2 冻存后间充质干细胞的活性率和增殖能力冻存可以减弱多发性骨髓瘤患者骨髓间充质干细胞的活性，并且与冻存时间有一定关系。短期冻存间充质干细胞的锥虫蓝拒染回收率为(92.9±7.5)%；长期冻存间充质干细胞的锥虫蓝拒染回收率为(86.7±9.2)%，二者相比差异有显著性意义(P < 0.05)。但是，冻存后间充质干细胞的增殖能力与冻存前比较，没有明显差异。依据生长曲线可以得出短期和长期冻存后骨髓间充质干细胞的倍增时间分别为(51.8±4.8) h和(49.6±3.6) h。 2.3 冻存后间充质干细胞抑制T淋巴细胞的增殖作用冻存后的多发性骨髓瘤患者骨髓间充质干细胞同样具有抑制T淋巴细胞增殖的作用，这种抑制作用随着间充质干细胞细胞数量的增加而增强。在间充质干细胞数量分别为0.5×104，1×104，2×104和5×104个时，相应的T淋巴细胞3H-TdT掺入的每分钟闪烁次数平均值分别为：短期冻存后(2.0±0.4)×104，(1.5±0.3)×104，(0.8±0.3)×104和(0.5±0.2)×104；长期冻存后(2.3±0.5)×104，(1.7±0.2)×104，(0.8±0.2)×104和(0.5±0.1)×104。而未加入间充质干细胞的CD3+T淋巴细胞每分钟闪烁次数平均值为(4.0±0.7)×104。冻存前多发性骨髓瘤患者骨髓间充质干细胞在上述细胞数量时的T淋巴细胞3H-TdT掺入的每分钟闪烁次数平均值分别为：(1.7±0.3)×104、(1.4± 0.4)×104、(0.9±0.2)×104和(0.3±0.1)×104。冻存前、后骨髓间充质干细胞抑制CD3+T淋巴细胞的增殖作用相似，见图1。"

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