Non-Enzymatic Generation of Placenta Single Cells From Third Trimester Human Placenta.

The placenta is a heterogeneous and complex organ with multiple cell types, posing a challenge for the maternal-fetal medicine field to implement single-cell technologies for a deeper characterization of this essential organ. Several protocols use enzymes to digest the tissue and generate single-cell suspensions, but this approach has several shortcomings, including the loss and reduced viability of cells. In this article, we describe a non-enzymatic approach to generate single-cell suspensions from placental tissue with high yield and viability for single-cell RNA sequencing. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Preparation of single-cell suspension from freshly collected third trimester human placental sections Basic Protocol 2: Visualization and determination of cell viability Alternate Protocol: Counting cells using a hemocytometer.