Isolation of Microparticles Derived from Apoptotic T Lymphocytes In Vitro

Published: August 31, 2023

Abstract

Source: Yang, C. et al., Generation of Lymphocytic Microparticles and Detection of their Proapoptotic Effect on Airway Epithelial Cells. J. Vis. Exp. (2015)

This video demonstrates the generation and isolation of T lymphocyte-derived microparticles, LMPs, from T lymphocytes following treatment with actinomycin-D.

Protocol

1. LMPs Production and Characterization NOTE: To prevent contamination, ensure that all materials used in this experiment are sterile or autoclaved. Perform all steps at RT in a biological safety cabinet under sterile conditions, unless otherwise indicated. Stimulation and Collection of MPs Thaw an aliquot of 10 million CEM T cells in a 37 °C water bath. Dilute in 10 ml pre-warmed serum-free hematopoietic medium such as X-VIVO…

Disclosures

The authors have nothing to disclose.

Materials

CEM T cells ATCC CCL-119
X-VIVO 15 medium Cambrex, Walkersville 04-744Q
Flask T75 Sarstedt 83.1813.502
Flask T175 Sarstedt 83.1812.502
Actinomycin D Sigma Chemical Co. A9415-2mg
PBS Lifetechnologies 14190-144
0.22µm filter Sarstedt 83.1826.001
Cell incubator Mandel Heracell 150
Low speed centrifuge IEC Centra8R
High speed centrifuge Beckman Avanti J8
High speed rotor for 250ml bottle Beckman JLA16.250
High speed rotor for 50ml tube Beckman JA30.50

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Cite This Article
Isolation of Microparticles Derived from Apoptotic T Lymphocytes In Vitro. J. Vis. Exp. (Pending Publication), e21570, doi: (2023).

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