2022 journal article

Fabrication and Use of Dry Macroporous Alginate Scaffolds for Viral Transduction of T Cells

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, (187).

MeSH headings : Alginates; Calcium; Immunotherapy, Adoptive; Receptors, Chimeric Antigen; T-Lymphocytes; Tissue Engineering; Tissue Scaffolds
TL;DR: Dry macroporous alginate scaffolds serve as a cheaper and more convenient alternative to the conventional transduction method for CAR-T cells and are comparable to the transduction efficiency of spinoculation on retronectin-coated plates. (via Semantic Scholar)
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Source: Web Of Science
Added: January 30, 2023

Genetic engineering of T cells for CAR-T cell therapy has come to the forefront of cancer treatment over the last few years. CAR-T cells are produced by viral gene transfer into T cells. The current gold standard of viral gene transfer involves spinoculation of retronectin-coated plates, which is expensive and time-consuming. There is a significant need for efficient and cost-effective methods to generate CAR-T cells. Described here is a method for fabricating inexpensive, dry macroporous alginate scaffolds, known as Drydux scaffolds, that efficiently promote viral transduction of activated T cells. The scaffolds are designed to be used in place of gold standard spinoculation of retronectin-coated plates seeded with virus and simplify the process for transducing cells. Alginate is cross-linked with calcium-D-gluconate and frozen overnight to create the scaffolds. The frozen scaffolds are freeze-dried in a lyophilizer for 72 h to complete the formation of the dry macroporous scaffolds. The scaffolds mediate viral gene transfer when virus and activated T cells are seeded together on top of the scaffold to produce genetically modified cells. The scaffolds produce >85% primary T cell transduction, which is comparable to the transduction efficiency of spinoculation on retronectin-coated plates. These results demonstrate that dry macroporous alginate scaffolds serve as a cheaper and more convenient alternative to the conventional transduction method.