Non-Enzymatic Generation of Placenta Single Cells from Third Trimester Human Placenta
Ameloko Joy, Idowu Aimola, Fomukong A Hanneda, Reuben B. Samson, Zeenat B Kudan, Habiba H. Abubakar, Musa Kana, Benedict Anchang
Abstract
The placenta is a heterogeneous and complex organ with multiple cell types, posing a challenge for the field of maternal-fetal medicine 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 suspension, but this approach has several shortcomings including the loss and reduced viability of cells. In this study, we describe a non-enzymatic approach to generate single cell suspension
from placental tissue with high yield and viability for single cell RNA
sequencing.
Before start
Attachments
non-enzymatic single cell generation edited.pdf
Steps
Placenta Single Cell Preparation Protocol
Transform placenta sections into gentle MACs C-tube containing 3000µL of MACs running buffer.
Set up a gentle MACS dissociator program to h-cord-01 for 553 rotation per round (rpr) at
0h 0m 30s .
After the run, centrifuge at 300x g,0h 0m 0s for 0h 10m 0s at 40°C.
Remove the supernatant leaving 200µL to resuspend pellet in. Transfer pipettes may be used
to remove supernatant after centrifugation to minimize loss of placenta cells.
10µL of placental cells can be counted on a countess II cell counter or using trypan blue.
