How to Isolate Human Regulatory T Cells (Tregs) Using the Easy 50 EasySep™ Magnet

This video is a step by step demonstration of how to isolate human regulatory T cells starting from up to 2 billion (2 x 10^9) peripheral blood mononuclear cells (PBMCs) or leukapheresis cells with the EasySep™ Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit (Catalog # 18063) and the Easy 50 EasySep™ Magnet (Catalog # 18002). Please also consult the Product Information Sheet for further details on the protocol.

EasySep™ Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit (Catalog# 18063): https://bit.ly/3aejV9v
Easy 50 EasySep™ Magnet (Catalog # 18002): https://bit.ly/3mLBra2
Product Information Sheet: https://bit.ly/2QqM9H3

00:00 Intro

04:04 Part 1: Isolation of CD25+ cells
04:15 Addition of the EasySep™ Human CD25 Positive Selection Cocktail (Catalog# 18063C.1)
06:15 Addition of the EasySep™ Releasable RapidSpheres™ 50201 and the EasySep™ Human CD4+ T Cell Enrichment Cocktail (Catalog# 19052C.2)
09:04 1st magnetic separation
11:52 Addition of EasySep™ Releasable RapidSpheres™ 50201 to supernatant collected after 1st magnetic separation
14:36 Transferring supernatant back to tube seated in magnet
15:13 2nd magnetic separation
16:26 Saving supernatant for optional isolation of CD4+CD25- responder T cells
16:37 3rd magnetic separation with wash step of CD25+ cells bound to EasySep™ Releasable RapidSpheres™ 50201
18:22 4th magnetic separation with wash step of CD25+ cells bound to EasySep™ Releasable RapidSpheres™ 50201
21:37 Sample resuspension + addition of Release Buffer

25:14 Part 2: Isolation of CD4+CD127low cells from CD25+ cell fraction
25:49 Addition of EasySep™ Human CD127high Depletion Cocktail (Catalog# 19233C)
27:05 Addition of the EasySep™ Dextran RapidSpheres™ 50100
29:06 Magnetic separation of the CD4+CD127lowCD25+ regulatory T cells

31:24 Optional Part: Isolation of CD4+CD25- responder T cells

#Treg #RegulatoryTcell #CellSeparation #CellIsolation #EasySep

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