Taggart M S, Tchir A, Van Dieren L, Chen H, Hassan M, Taveras C, Lellouch A G, Toner M, Sandlin R D, Uygun K
Center for Engineering in Medicine and Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA.
Shriners Children's Boston, Boston, MA, USA.
bioRxiv. 2024 Jul 17:2024.07.14.603471. doi: 10.1101/2024.07.14.603471.
Drug discovery pipelines rely on the availability of isolated primary hepatocytes for investigating potential hepatotoxicity prior to clinical application. These hepatocytes are typically isolated from livers rejected for transplantation and subsequently cryopreserved for later usage. The gold-standard cryopreservation technique, slow-freezing, is a labor-intensive process, with significant post-storage viability loss. In this work, we introduce parallelized droplet vitrification, a technique for high-volumetric, rapid vitrification of suspended cells. We show the utility of this technique through the single-run vitrification of the whole-rate liver hepatocyte yield, resulting in a 1600% increase in single-batch vitrification and a 500% increase in droplet generation rate compared to previous droplet vitrification approaches. Additionally, we showed that these implementations maintained improved post-preservation outcomes in primary rat hepatocytes.
药物研发流程依赖于分离的原代肝细胞，以便在临床应用前研究潜在的肝毒性。这些肝细胞通常从被拒绝用于移植的肝脏中分离出来，随后进行冷冻保存以备后用。金标准冷冻保存技术——慢速冷冻，是一个劳动密集型过程，储存后细胞活力会显著丧失。在这项工作中，我们引入了并行化液滴玻璃化技术，这是一种用于悬浮细胞的大容量、快速玻璃化技术。我们通过对全量肝脏肝细胞产量进行单次玻璃化展示了该技术的实用性，与之前的液滴玻璃化方法相比，单批次玻璃化量增加了1600%，液滴生成速率提高了500%。此外，我们还表明，这些方法在原代大鼠肝细胞中保持了更好的保存后效果。
