Isolation of Mononuclear Cells from Peripheral Blood and Decidual Tissue

Peripheral blood samples were layered on Ficoll Hypaque gradients (LymphoprepTM; Alere Technologies, Norway) for density gradient centrifugation (453 × g for 30 min). Mononuclear cells were isolated and washed twice with phosphate-buffered saline (PBS). Decidual tissues were rinsed thoroughly with PBS and minced into 1–2 mm pieces by a pair of scalpel blades in Dulbecco's Modified Eagle Medium. Then, the suspensions were filtered through a 32 μm nylon mesh as reported elsewhere (24 (link)). All samples were cryopreserved until single-cell analysis.

Free full text: Click here

Morita K., Tsuda S., Kobayashi E., Hamana H., Tsuda K., Shima T., Nakashima A., Ushijima A., Kishi H, & Saito S. (2020). Analysis of TCR Repertoire and PD-1 Expression in Decidual and Peripheral CD8+ T Cells Reveals Distinct Immune Mechanisms in Miscarriage and Preeclampsia. Frontiers in Immunology, 11, 1082.

Publication 2020

LymphoprepTM;

Blood Cells Decidual Density gradient centrifugation Eagle Ficoll Hypaque Nylon Phosphate Saline Single cell analysis Tissues

Corresponding Organization : University of Toyama

Top 5 similar protocols

Variable analysis

independent variables

Density gradient centrifugation (453 × g for 30 min)
Mincing of decidual tissues into 1–2 mm pieces
Filtering of suspensions through a 32 μm nylon mesh

dependent variables

Isolation of mononuclear cells

control variables

Phosphate-buffered saline (PBS) for washing mononuclear cells
Dulbecco's Modified Eagle Medium for mincing decidual tissues

controls

No positive or negative controls were explicitly mentioned in the input protocol.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!