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A pair of cell preservation solutions for therapy with human adipose tissue-derived mesenchymal stromal cells.
Regenerative Therapy ( IF 4.3 ) Pub Date : 2020-01-16 , DOI: 10.1016/j.reth.2019.10.004
Yasutaka Fujita 1 , Masuhiro Nishimura 1 , Natsuki Watanabe Komori 1 , Tamaki Wada 1 , Chikage Shirakawa 1 , Taichi Takenawa 1 , Osamu Sawamoto 1 , Masako Doi 1
Affiliation  

Introduction

Stem cells for therapy are often suspended in a preservation solution, such as normal saline or lactated Ringer's solution, for a short time before intravenous infusion. However, these solutions are not necessarily ideal for maintaining cell viability and preventing the sedimentation of cells during storage and infusion. In this study, we attempted to optimize the compositions of preservation solutions, which could affect the efficacy and safety of stem cell therapy.

Methods

We determined the characteristics of a preservation solution that would optimize cell viability and the percentage of cells in the supernatant using human adipose-derived mesenchymal stromal cells (hADSCs). We compared solutions that differed by electrolytes (e.g., normal saline and Ringer's solution) and the concentrations of dextran 40 and trehalose. The effects of the solutions on hADSCs were evaluated by assessing cell surface markers, colony-forming capacity, differentiation potential, and cell concentrations in the infusion line.

Results

Optimized preservation solutions consisted of lactated Ringer's solution with 3% trehalose without or with 5% dextran 40 (LR-3T and LR-3T-5D, respectively). The cell viabilities after 24 h of storage at 5 °C in LR-3T and LR-3T-5D were 94.9% ± 2.4% and 97.6% ± 2.4%, respectively. The percentage of cells in the supernatant after 1 h of storage at room temperature in LR-3T-5D was 83.5% ± 7.6%. These solutions preserved the percentage of cell surface marker-positive cells, the colony-forming capacity, and the adipogenic and osteogenic differentiation ability in hADSCs for at least 24 h after preservation at 5 °C and 25 °C.

Discussion

We determined the optimal composition of preservation solutions for hADSCs and confirmed the effects of these solutions on cell viability and the stability of cell characteristics in vitro. Our results suggest that LR-3T and LR-3T-5D can help maintain the quality of stem cells for therapy during preservation and infusion. However, further in vivo research is needed on the efficacy and safety of the solutions in different therapeutic cell lines before clinical use.



中文翻译:

用于治疗人类脂肪组织来源的间充质基质细胞的一对细胞保存溶液。

介绍

用于治疗的干细胞通常在静脉输注前短时间悬浮在保存溶液中,例如生理盐水或乳酸林格溶液。然而,这些解决方案对于维持细胞活力和防止细胞在储存和输注过程中沉降并不一定是理想的。在这项研究中,我们试图优化保存液的成分,这可能会影响干细胞治疗的疗效和安全性。

方法

我们使用人脂肪来源的间充质基质细胞 (hADSC) 确定了可以优化细胞活力和上清液中细胞百分比的保存溶液的特性。我们比较了因电解质(例如生理盐水和林格溶液)以及葡聚糖 40 和海藻糖浓度而异的溶液。通过评估细胞表面标志物、集落形成能力、分化潜能和输注线中的细胞浓度来评估溶液对 hADSCs 的影响。

结果

优化的保存溶液由乳酸林格溶液和 3% 海藻糖组成,不含或含 5% 葡聚糖 40(分别为 LR-3T 和 LR-3T-5D)。LR-3T 和 LR-3T-5D 在 5°C 下储存 24 小时后的细胞活力分别为 94.9% ± 2.4% 和 97.6% ± 2.4%。LR-3T-5D在室温下储存1小时后上清液中的细胞百分比为83.5%±7.6%。这些溶液在 5°C 和 25°C 保存后至少 24 小时保留了 hADSCs 中细胞表面标记阳性细胞的百分比、集落形成能力以及脂肪形成和成骨分化能力。

讨论

我们确定了hADSCs保存液的最佳组成,并证实了这些溶液对细胞活力和体外细胞特性稳定性的影响。我们的研究结果表明,LR-3T 和 LR-3T-5D 有助于在保存和输注过程中维持干细胞的质量以进行治疗。然而,在临床使用前,还需要进一步研究溶液在不同治疗细胞系中的功效和安全性

更新日期:2020-01-16
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