Method for preparing CAR-T cell by CRISPR/Cas9
A cell and DNA sequence technology, applied in the field of biology, to achieve the effect of high editing efficiency, avoiding potential safety hazards and high safety
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0020] Example 1 (Preparation Example)
[0021] Use CRISRP / Cas9 system to produce leukemia CAR-T cells:
[0022] 1) T cell separation and culture
[0023] a. Separate fresh peripheral blood mononuclear cells (PBMC, peripheral blood mononuclear cell) by density gradient centrifugation;
[0024] b. Use paramagnetic beads coupled with anti-CD3 and anti-CD28 antibodies (Dynabeads ClinExVivo CD3 / CD28, Invitrogen, Camarillo, CA, USA) to enrich CD3+ cells, the ratio of magnetic beads to cells is 3:1; the cells are diluted to TNC (Total nucleated cell) concentration is 20-30x106 / mL, incubated with magnetic beads in a petri dish for 2 hours at room temperature;
[0025] c. Use Magnetic particles concentrator (MPC) (Invitrogen) to enrich CD3+ cells; cells containing CD3+ are resuspended in culture medium (OpTmizer TM CTS TM In T-Cell Expansion SFM, Life Technologies), the final concentration is 1x 106 cells / mL. Incubate for 2 days in a 37°C, 5% CO2 incubator.
[0026] 2) Construction of CRIS...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap