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Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood

Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood

DOI:
10.3791/50070-v

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09:29 min

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February 01, 2013

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, , , , , , , ,
1Division of Pediatrics,U.T. MD Anderson Cancer Center, 2Department of Stem Cell Transplantation and Cellular Therapy,U.T. MD Anderson Cancer Center

Chapters

  • 00:05Title
  • 02:09Isolation of Mononuclear Cells from PB and UCB
  • 03:04Preparation of T Cells for Electroporation
  • 04:03Electroporation of MNC
  • 05:59FACS Analysis of CAR Expression (Day 1)
  • 06:32Preparation of aAPC (Clone #4) on Day 1
  • 07:01aAPC-mediated Stimulation of CAR+ T Cells (Day 1)
  • 07:33Results: SB System Coupled with Co-culture on aAPC Generates CD19-specific CAR+ T Cells
  • 08:44Conclusion

Summary

Automatic Translation

Automatic Translation

T cells expressing a CD19-specific chimeric antigen receptor (CAR) are infused as investigational treatment of B-cell malignancies in our first-in-human gene therapy trials. We describe genetic modification of T cells using the Sleeping Beauty (SB) system to introduce CD19-specific CAR and selective propagation on designer CD19+ artificial antigen presenting cells.

Tags

Keywords: Sleeping BeautyArtificial Antigen Presenting CellsGenetic ModificationT CellsPeripheral BloodUmbilical Cord BloodCAR T CellsCD19Gene TherapyImmunotherapyTumor-associated AntigensClinical Application

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