Center for Reproductive Health Sciences, Department of Obstetrics and Gynecology, Washington University School of Medicine, St Louis, MO, USA.
Division of Obstetrics and Gynecology, School of Medicine, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
Methods Mol Biol. 2024;2781:143-154. doi: 10.1007/978-1-0716-3746-3_13.
The dissociation of whole tissue into single-cell suspensions is a critical step for techniques focused on profiling of individual cells. Here, we describe a protocol for the preparation of high-quality single-cell suspensions from human placental tissues: the basal plate (BP), placental villi (PV), and chorioamniotic membranes (CAM). This protocol also provides guidance for the cryopreservation and recovery of single-cell suspensions for later use. The methods described here have been demonstrated to be suitable for downstream single-cell applications, such as single-cell RNA-sequencing, that require viable, high-quality cell suspensions.
将整个组织解离成单细胞悬液是专注于单个细胞分析的技术的关键步骤。在这里,我们描述了一种从人胎盘组织中制备高质量单细胞悬液的方案:基底板(BP)、胎盘绒毛(PV)和羊膜绒毛膜(CAM)。该方案还为单细胞悬液的冷冻保存和恢复提供了指导,以便以后使用。这里描述的方法已被证明适用于需要有活力、高质量细胞悬液的单细胞 RNA 测序等下游单细胞应用。