Development of Serum-Free Media for Cryopreservation of Hydrogel Encapsulated Cell-Based Therapeutics
Development of Serum-Free Media for Cryopreservation of Hydrogel Encapsulated Cell-Based Therapeutics
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开发用于水凝胶封装的细胞疗法冷冻保存的无血清培养基
DOI：
10.1007/s12195-022-00739-7
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发表时间：
2022
影响因子：
2.8
通讯作者：
Veiseh, Omid
中科院分区：
文献类型：
--
作者：
Cui, Yufei;Nash, Amanda M.;Castillo, Bertha;Sanchez Solis, Leonardo D.;Aghlara-Fotovat, Samira;Levitan, Maya;Kim, Boram;Diehl, Michael;Veiseh, Omid
虽然水凝胶包封细胞已被开发用于治疗多种疾病，但仍需要冷冻保存和维持治疗性包封细胞产品的复合功能的方法以促进其储存和分布。虽然方法来保存封装的细胞，和合成后已经得到了最近的关注，有效的保存介质还没有完全defined.MethodsWe采用了一个两层筛选的初始库的32个不同的冷冻保存剂（CPA）配方组成的不同的细胞渗透和不渗透剂。配方进行了测定，使用暗视野显微镜，以评估藻酸盐水凝胶基质的完整性，然后通过细胞活力分析和功能分泌activity.ResultsThe结构完整性的大> 1毫米藻酸盐胶囊是高度敏感的冷冻和解冻在单独的媒体，但可以恢复由一些CPA配方含有不同的细胞渗透和渗透剂。随后的活力筛选确定了两种性能最佳的CPA制剂，其在-80 °C储存后最大化了胶囊完整性和细胞活力。顶部配方（10%二甲基亚砜（DMSO）和0.3 M葡萄糖）被证明，以保持水凝胶的完整性和保留细胞活力超过一个关键的美国FDA设置70%的活力阈值，同时保持蛋白质分泌和所得的细胞potenti.ConclusionsThis优先筛选确定了冷冻保存解决方案，保持大海藻酸盐胶囊的完整性，并产生高的活力和效力。重要的是，这种制剂是无血清的，无毒的，并且可以支持临床上可翻译的基于包封细胞的治疗剂的开发。
IntroductionWhile hydrogel encapsulation of cells has been developed to treat multiple diseases, methods to cryopreserve and maintain the composite function of therapeutic encapsulated cell products are still needed to facilitate their storage and distribution. While methods to preserve encapsulated cells, and post-synthesis have received recent attention, effective preservation mediums have not been fully defined.MethodsWe employed a two-tiered screen of an initial library of 32 different cryopreservation agent (CPA) formulations composed of different cell-permeable and impermeable agents. Formulations were assayed using dark field microscopy to evaluate alginate hydrogel matrix integrity, followed by cell viability analyses and measurements of functional secretion activity.ResultsThe structural integrity of large > 1 mm alginate capsules were highly sensitive to freezing and thawing in media alone but could be recovered by a number of CPA formulations containing different cell-permeable and impermeable agents. Subsequent viability screens identified two top-performing CPA formulations that maximized capsule integrity and cell viability after storage at − 80 °C. The top formulation (10% Dimethyl sulfoxide (DMSO) and 0.3 M glucose) was demonstrated to preserve hydrogel integrity and retain cell viability beyond a critical USA FDA set 70% viability threshold while maintaining protein secretion and resultant cell potency.ConclusionsThis prioritized screen identified a cryopreservation solution that maintains the integrity of large alginate capsules and yields high viabilities and potency. Importantly, this formulation is serum-free, non-toxic, and can support the development of clinically translatable encapsulated cell-based therapeutics.
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DOI：
10.1002/advs.202002425
发表时间：
2021-03
期刊：
Advanced science (Weinheim, Baden-Wurttemberg, Germany)
影响因子：
--
作者：
Chang T;Zhao G
通讯作者：
Zhao G
细胞封装技术作为中枢神经系统恶性肿瘤的治疗策略。
影响因子：
15.9
作者：
T. Visted;R. Bjerkvig;P. Enger
通讯作者：
P. Enger
DOI：
10.1007/978-1-62703-550-7_23
发表时间：
2013-01-01
期刊：
IMMOBILIZATION OF ENZYMES AND CELLS, 3RD EDITION
影响因子：
--
作者：
Acarregui, Argia;Orive, Gorka;Maria Hernandez, Rosa
通讯作者：
Maria Hernandez, Rosa
影响因子：
3
作者：
Chen, Bo;Wright, Bernice;Connon, Che J.
通讯作者：
Connon, Che J.
影响因子：
2.7
作者：
Petrenko, Yu. A.;Jones, D. R. E.;Petrenko, A. Yu.
通讯作者：
Petrenko, A. Yu.