Generation of an In Vitro Cell Culture Model of Malaria-HIV Co-Infection

Published: October 31, 2023

Abstract

Source: Finney, C. et al., An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection. J. Vis. Exp. (2015)

This video demonstrates the generation of an in vitro malaria-HIV co-infection cell culture model. HIV-infected peripheral blood mononuclear cells co-cultured with Plasmodium falciparum parasitized erythrocytes potentially support each other's growth and provide insights into the immune responses and disease progression in HIV and malaria co-infections.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. CAUTION: Working with human blood samples and human malaria parasites requires precautionary measures. Always wear a lab coat, and gloves, and work in a level 2 Biosafety cabinet. In the event of accidental percutaneous exposure to human malaria, report to Health and Safety for prophylactic treatment. Additional safety considerations should also be put in place…

Representative Results

Figure 1. Depiction of Ficoll gradient post spin demonstrating the position of the PBMCs.

開示

The authors have nothing to disclose.

Materials

Alanine Sigma A7377
BD Vacutainer ACD Solution A BD 364506
BD Vacutainer Sodium Heparin BD 17-1440-02
DPBS (no calcium, no magnesium) Corning 21-031-CV
Fetal Bovine Serum Sigma F1051 Heat inactivate before use
Ficoll-Paque PLUS GE Healthcare 17-1440-02
Gentamycin (10mg/ml) Gibco 15710-064
Hema3 Staining Set Fisher 122-911
HEPES Fisher BP310-500
Hypoxanthine Sigma H9636
Ionomycin Sigma I3909
MEM non-essential amino acids (10mM) Gibco 11140
PMA Sigma P8139
RPMI-1640 powder Life-Technologies 31800-022
RPMI-1640 with L-glutamine and HEPES Thermo Scientific SH30255.01
Sodium bicarbonate (powder, cell culture) Sigma S5761
Sodium Pyruvate (100mM) Gibco 11360
Tris (Trizma base) Sigma T6066
Trizol Ambion 15596018
Trypan Blue (0.4%) Gibco 15250-061
Parasite Gas Mixture By special order 3% CO2, 1% O2, balance N2

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記事を引用
Generation of an In Vitro Cell Culture Model of Malaria-HIV Co-Infection. J. Vis. Exp. (Pending Publication), e21717, doi: (2023).

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