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间充质基质细胞的冷冻保存与复苏

Cryopreservation and revival of mesenchymal stromal cells.

作者信息

Haack-Sørensen Mandana, Kastrup Jens

机构信息

Cardiology Stem Cell Laboratory and Cardiac Catheterization Laboratory, The Heart Centre, Rigshospitalet Copenhagen University Hospital, Copenhagen, Denmark.

出版信息

Methods Mol Biol. 2011;698:161-74. doi: 10.1007/978-1-60761-999-4_13.

Abstract

Over the past few years, the pace of preclinical stem cell research is astonishing and adult stem cells have become the subject of intense research. Due to the presence of promising supporting preclinical data, human clinical trials for stem cell regenerative treatment of various diseases have been initiated. As there has been a precedent for the use of bone marrow stem cells in the treatment of hematological malignancies and ischemic heart diseases through randomized clinical safety and efficacy trials, the development of new therapies based on culture-expanded human mesenchymal stromal cells (MSCs) opens up new possibilities for cell therapy. To facilitate these applications, cryopreservation and long-term storage of MSCs becomes an absolute necessity. As a result, optimization of this cryopreservation protocol is absolutely critical. The major challenge during cellular cryopreservation is the lethality associated with the cooling and thawing processes. The major objective is to minimize damage to cells during low temperature freezing and storage and the use of a suitable cryoprotectant. The detrimental effects of cellular cryopreservation can be minimized by controlling the cooling rate, using better cryoprotective agents, maintaining appropriate storage temperatures, and controlling the cell thawing rate. As is described in this chapter, human MSCs can either be frozen in cryovials or in freezing bags together with cryopreserve solutions containing dimethyl sulfoxide (DMSO).

摘要

在过去几年中,临床前干细胞研究的步伐惊人,成体干细胞已成为深入研究的对象。由于存在有前景的临床前支持数据,针对各种疾病的干细胞再生治疗的人体临床试验已经启动。鉴于通过随机临床安全性和有效性试验使用骨髓干细胞治疗血液系统恶性肿瘤和缺血性心脏病已有先例,基于培养扩增的人间充质基质细胞(MSC)开发新疗法为细胞治疗开辟了新的可能性。为了促进这些应用,MSC的冷冻保存和长期储存成为绝对必要。因此,优化这种冷冻保存方案至关重要。细胞冷冻保存期间的主要挑战是与冷却和解冻过程相关的致死性。主要目标是在低温冷冻和储存期间将对细胞的损伤降至最低,并使用合适的冷冻保护剂。通过控制冷却速率、使用更好的冷冻保护剂、维持适当的储存温度以及控制细胞解冻速率,可以将细胞冷冻保存的有害影响降至最低。如本章所述,人MSC可以与含有二甲基亚砜(DMSO)的冷冻保存溶液一起冷冻在冻存管或冷冻袋中。

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