Cryopreservation with a twist - Towards a sterile, serum-free surface-based vitrification of hESCs
Cryopreservation with a twist - Towards a sterile, serum-free surface-based vitrification of hESCs
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DOI：
10.1016/j.cryobiol.2012.10.001
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发表时间：
2013-02-01
期刊：
影响因子：
2.7
通讯作者：
Zimmermann, H.
中科院分区：
文献类型：
--
作者：
Beier, A. F.;Schulz, J. D.;Zimmermann, H.
人类胚胎干细胞在再生医学、基础科研、组织工程和毒理学等领域发挥着重要作用。然而，它们独特的形态使它们对冷冻保存程序非常敏感。我们最近引入了一种表面依赖性、无酶和无血清的方法，用于使用直接浸入液氮中有效冷冻保存大量hESC集落（Beier等人，2011 [5]）。然而，直接接触液氮的风险污染和细胞感染，严重限制了临床应用，这项工作介绍了一种改进的方法和一种新的组合培养和冷冻保存装置，以促进表面依赖性玻璃化，而不接触（可能未灭菌）液氮。该技术允许在同一装置中进行培养、冷冻保存、储存和解冻后培养，而无需将细胞样品从培养表面分离。在没有直接液氮接触的情况下成功玻璃化大量hESC是朝向用于干细胞和其他集落形成细胞类型的临床应用的自动化冷冻保存过程的重要一步。(c)2012 Elsevier Inc. All rights reserved.
Human embryonic stem cells (hESCs) play an important role in the fields of regenerative medicine, basic scientific research, tissue engineering and toxicology. Their unique morphology however makes them very sensitive to cryopreservation procedures. We recently introduced a surface dependent, enzyme- and serum-free method for the effective cryopreservation of bulk quantities of hESC colonies using direct immersion into liquid nitrogen (Beier et al., 2011 [5]). However, direct contact with liquid nitrogen risks contamination and cell infection and severely limits clinical application.This work introduces a modified method and a new combined cultivation and cryopreservation device to facilitate the surface dependent vitrification without contact with (possibly unsterile) liquid nitrogen. The technique allows the culture, cryopreservation, storage and post-thawing cultivation in the same device without detaching cell samples from the cultivation surface. Successful vitrification of bulk quantities of hESCs without direct liquid nitrogen contact is an important step towards automated cryopreservation processes for clinical applications of stem cells and other colony forming cell types. (c) 2012 Elsevier Inc. All rights reserved.