Petrenko Yuriy, Chudickova Milada, Vackova Irena, Groh Tomas, Kosnarova Eliska, Cejkova Jitka, Turnovcova Karolina, Petrenko Alexander, Sykova Eva, Kubinova Sarka
Institute of Experimental Medicine of the Czech Academy of Sciences, Prague, Czech Republic.
Bioinova Ltd., Prague, Czech Republic.
Stem Cells Int. 2019 Jan 20;2019:5909524. doi: 10.1155/2019/5909524. eCollection 2019.
The wide use of human multipotent mesenchymal stromal cells (MSCs) in clinical trials requires a full-scale safety and identity evaluation of the cellular product and subsequent transportation between research/medical centres. This necessitates the prolonged hypothermic storage of cells prior to application. The development of new, nontoxic, and efficient media, providing high viability and well-preserved therapeutic properties of MSCs during hypothermic storage, is highly relevant for a successful clinical outcome. In this study, a simple and effective trehalose-based solution was developed for the hypothermic storage of human bone marrow MSC suspensions for further clinical applications. Human bone marrow MSCs were stored at 4°C for 24, 48, and 72 hrs in the developed buffered trehalose solution and compared to several research and clinical grade media: Plasma-Lyte® 148, HypoThermosol® FRS, and Ringer's solution. After the storage, the preservation of viability, identity, and therapeutically associated properties of MSCs were assessed. The hypothermic storage of MSCs in the new buffered trehalose solution provided significantly higher MSC recovery rates and ability of cells for attachment and further proliferation, compared to Plasma-Lyte® 148 and Ringer's solution, and was comparable to research-grade HypoThermosol® FRS. There were no differences in the immunophenotype, osteogenic, and adipogenic differentiation and the immunomodulatory properties of MSCs after 72 hrs of cold storage in these solutions. The obtained results together with the confirmed therapeutic properties of trehalose previously described provide sufficient evidence that the developed trehalose medium can be applied as a low-cost and efficient solution for the hypothermic storage of MSC suspensions, with a high potential for translation into clinical practice.
人多能间充质基质细胞(MSCs)在临床试验中的广泛应用,需要对细胞产品进行全面的安全性和身份评估,并在研究/医疗中心之间进行后续运输。这就需要在应用前对细胞进行长时间的低温保存。开发新的、无毒且高效的培养基,以在低温保存期间保持MSCs的高活力和良好保存的治疗特性,对于成功的临床结果至关重要。在本研究中,开发了一种简单有效的基于海藻糖的溶液,用于人骨髓MSC悬浮液的低温保存,以供进一步临床应用。将人骨髓MSCs在开发的缓冲海藻糖溶液中于4°C保存24、48和72小时,并与几种研究和临床级培养基进行比较:Plasma-Lyte® 148、HypoThermosol® FRS和林格氏液。保存后,评估MSCs的活力、身份和治疗相关特性的保存情况。与Plasma-Lyte® 148和林格氏液相比,在新的缓冲海藻糖溶液中低温保存MSCs可提供显著更高的MSC回收率以及细胞附着和进一步增殖的能力,并且与研究级HypoThermosol® FRS相当。在这些溶液中冷藏72小时后,MSCs的免疫表型、成骨和成脂分化以及免疫调节特性没有差异。所获得的结果以及先前描述的海藻糖已证实的治疗特性提供了充分的证据,表明所开发的海藻糖培养基可作为一种低成本且高效的溶液用于MSC悬浮液的低温保存,具有很高的转化为临床实践的潜力。