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 |
