人脐带间充质干细胞新型冻存液实验研究

张鹏; 刘宝林

doi:10.12465/j.issn.0253-4339.2025.03.158

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人脐带间充质干细胞新型冻存液实验研究

更新时间：2025-05-30

- 人脐带间充质干细胞新型冻存液实验研究
- Experimental Study on a Novel Solution for Cryopreservation of Human Umbilical Cord Mesenchymal Stem Cells
- 制冷学报 2025年46卷第3期页码：158-166
- 作者机构：
  
  上海理工大学生物系统热科学研究所　上海　 200093
- 作者简介：
  
  刘宝林，男，教授，上海理工大学生物系统热科学研究所，13636524955，E-mail：blliuk@163.com。研究方向：低温生物医学工程。
- 基金信息：
  
  国家自然科学基金(51776130)
- DOI：10.12465/j.issn.0253-4339.2025.03.158
  中图分类号： TB61⁺1;R318.52
- 收稿日期：2024-02-27，
  
  修回日期：2024-03-24，
  
  录用日期：2024-04-10，
  
  纸质出版日期：2025-06-16
- 稿件说明：
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张鹏, 刘宝林. 人脐带间充质干细胞新型冻存液实验研究[J]. 制冷学报, 2025,46(3):158-166.

Zhang Peng, Liu Baolin. Experimental Study on a Novel Solution for Cryopreservation of Human Umbilical Cord Mesenchymal Stem Cells[J]. Journal of refrigeration, 2025, 46(3): 158-166.
张鹏, 刘宝林. 人脐带间充质干细胞新型冻存液实验研究[J]. 制冷学报, 2025,46(3):158-166. DOI： 10.12465/j.issn.0253-4339.2025.03.158.

Zhang Peng, Liu Baolin. Experimental Study on a Novel Solution for Cryopreservation of Human Umbilical Cord Mesenchymal Stem Cells[J]. Journal of refrigeration, 2025, 46(3): 158-166. DOI： 10.12465/j.issn.0253-4339.2025.03.158.

摘要

细胞库普遍使用的二甲基亚砜（Me

SO）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Me

SO并具有与Me

SO产品相当的效果。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+10 g/L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（体积分数为10%Me

SO）无显著性差异。冻存细胞培养3 d后细胞数量为（12.42±0.60）× 10

个（增殖倍数为4.97），且细胞表型与新鲜细胞无显著性差异。新型干细胞冻存液解决了细胞低温保存中的“Me

SO-free”问题，具有潜在的应用前景。

Abstract

Dimethyl sulfoxide (Me

SO) in cell banking exhibits significant side effects on both the cells and the human body. Therefore

an approach that mitigates the side effects of Me

SO with comparable efficacy is urgently needed. The human umbilical cord mesenchymal was used as the research material. First

the thermal physical properties of trehalose

glucose

and L-proline and their regulation of ice crystal growth were measured using a differential scanning calorimeter and a cryomicroscope. Cryopreservation experiments were performed to determine the optimal concentration of each component in the cryopreservation solution

and the viability and functionality of the cells after cryopreservation were validated. The 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+10 g/L whey protein+0.1 mol/L trehalose+Normosol-R) and the conventional cryopreservation solution (a volume fraction of 10% Me

SO). Moreover

after 3 days of culture

the cell number was (12.42±0.60) × 10

(proliferation fold of 4.97)

and the cell phenotype was not significantly different from that of fresh cells. The proposed novel solution for stem cell cryopreservation solves the problem of "Me

SO-free" cryopreservation of cells and offers promising potential for clinical applications.

关键词

Keywords

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