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人脐带间充质干细胞新型冻存液实验研究
张鹏, 刘宝林
0
(生物系统热科学研究所 上海)
摘要:
细胞库普遍使用的二甲基亚砜(Me2SO)对细胞和人体均有较强的副作用,因此亟需一种方案来替代Me2SO并具有与Me2SO产品相当效果。实验以人脐带间充质干细胞为研究材料,首先通过差示扫描量热仪和低温显微系统测定了海藻糖、葡萄糖和L-脯氨酸的热物性及其对冰晶生长的调控,在此基础上通过冻存实验确定了冻存液中各组分的浓度,并验证了冻存后的细胞的活性和功能。结果显示,使用新型干细胞冻存液(1.25 mol/L乙二醇 + 1.0%乳清蛋白 + 0.1 mol/L海藻糖 + Normosol-R注射液)冻存后的细胞存活率(92.42 ± 0.28%)和回收率(87.80 ± 4.22%)与传统冻存液(10%Me2SO)无显著性差异,且新型干细胞冻存液能够更好地维持干细胞的增殖能力和表型。新型干细胞冻存液解决了细胞低温保存中的“Me2SO-free”问题,具有潜在的应用前景。
关键词:  低温保存  细胞治疗  干细胞  无Me2SO  细胞冻存液
DOI:
投稿时间:2024-02-27  修订日期:2024-03-24   录用日期:2024-04-10
基金项目:国家自然科学基金(三维多孔微载体/肝细胞复合体低温保存的关键问题)
Experimental Study on a Novel Cryopreservation Solution for Human Umbilical Cord Mesenchymal Stem Cells
Abstract:
The commonly used dimethyl sulfoxide (Me2SO) in cell banking exhibits significant side effects on both cells and the human body. Therefore, there is an urgent need for an approach to mitigate the side effects of Me2SO while with comparable efficacy. The study uses human umbilical cord mesenchymal stem cells as the research material. First, the thermal physical properties of trehalose, glucose and L-proline and their regulation of ice crystal growth were measured by a differential scanning calorimeter and a cryomicroscope. Further, cryopreservation experiments were performed to determine the optimal concentrations of each component in the cryopservation solution, and the viability and functionality of cells after cryopreservation were validated. Results show that there is no significant difference in cell viability (92.42 ± 0.28%) and recovery rate (87.80 ± 4.22%) between the use of the novel stem cell cryopreservation solution (1.25 mol/L ethylene glycol + 1.0% whey protein + 0.1 mol/L trehalose + Normosol-R) and conventional cryopreservation solution (10%Me2SO). Moreover, the novel stem cell cryopservation solution better preserves the proliferative capacity and stemness of stem cells. The proposed novel stem cell cryopservation solution addresses the "Me2SO-free" issue in cell cryopreservation, holding promising potential for clinical applications.
Key words:  Cryopreservation  Cell therapy  Mesenchymal stem cells  Me2SO-free  Cryopreservation solution

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