Comparison of three methodologies of isolation, purification and in vitro culture of murine T lymphocytes
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| Auteurs: | , , , , , |
|---|---|
| Format: | artículo original |
| Statut: | Versión publicada |
| Date de publication: | 2019 |
| Description: | The isolation and selection of pure T lymphocytes from spleen primary culture can be obtained easily and quickly using commercial kits that use magnetics microbeads to capture and purify the cells. Despite these advantages, the financial burden that the constant acquisition of these kits can impose to a laboratory budget makes it important to evaluate other options. We compared the cell recovery, viability and purity percentage of three isolation and purification methods; two of them of low cost and easy implementation (cell adhesion of cultures treated with Ammonium-Chloride-Potassium (ACK) buffer, and Ficoll), and a commercial with kit of negative selection with magnetics microbeads. With the ACK-cell adhesion method, we found a recovery and purity percentage of lymphocytes T comparable to the one obtained with the kit. Although this method requires more time, as cells must be incubated overnight, it is easily implemented and has a comparable efficiency to the kit, as well as a lower cost. This method can be taken into account for experiments that can use activated cells. |
| Pays: | Portal de Revistas TEC |
| Institution: | Instituto Tecnológico de Costa Rica |
| Repositorio: | Portal de Revistas TEC |
| Langue: | Español |
| OAI Identifier: | oai:ojs.pkp.sfu.ca:article/4352 |
| Accès en ligne: | https://revistas.tec.ac.cr/index.php/tec_marcha/article/view/4352 |
| Mots-clés: | CD3 ACK buffer cellular adherence Ficoll magnetic microbeads amortiguador ACK adherencia celular microesferas magnéticas |