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Related Concept Videos

Tumor Immunotherapy01:27

Tumor Immunotherapy

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Immunotherapy is a treatment that boosts or manipulates the immune system to fight diseases, including cancer. For instance, by stimulating an immune response through vaccinations against viruses that cause cancers, like hepatitis B virus and human papillomavirus, these diseases can be prevented. Nonetheless, some cancer cells can avoid the immune system due to their rapid mutation and division. The immune response to many cancers involves three phases: elimination, equilibrium, and escape.
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Cancer Cell Migration through Invadopodia01:35

Cancer Cell Migration through Invadopodia

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Invadosome is a broad category of cell surface structures with proteolytic activity that  degrades the extracellular matrix (ECM). Invadosomes are present in normal cell types, including macrophages, endothelial cells, and neurons, as well as tumor cells. Although the macrophage podosomes and tumor cell invadopodia are classified as invadosomes, they have different structures, molecular pathways, and functions. Podosomes are short structures that last for a few minutes. However,...
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Adaptive Mechanisms in Cancer Cells02:53

Adaptive Mechanisms in Cancer Cells

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Cancer cells accumulate genetic changes at an abnormally rapid rate due to the defects in the DNA repair mechanisms. From an evolutionary perspective, such genetic instability is advantageous for cancer development. Mutant cell lines accumulate a series of beneficial mutations that contribute to their progression into cancer.
Some of the advantages that cancer cells have on normal cells include - enhanced ability to divide without terminally differentiating, induce new blood vessel formation,...
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Cancer Stem Cells and Tumor Maintenance02:40

Cancer Stem Cells and Tumor Maintenance

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Early diagnosis and treatment can often cure cancer. However, even with treatment, residual cells called cancer stem cells (CSC) might remain, often causing tumor recurrence. These cancer stem cells possess the potential for self-renewal and multi-lineage differentiation and are often responsible for the therapeutic resistance displayed in most cancers.
Cancer stem cells are thought to originate from tissue-specific normal stem cells or progenitor cells. The normal stem cells usually reside in...
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Cancers Originate from Somatic Mutations in a Single Cell02:21

Cancers Originate from Somatic Mutations in a Single Cell

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Cancer arises from mutations in the critical genes that allow healthy cells to escape cell cycle regulation and acquire the ability to proliferate indefinitely. Though originating from a single mutation event in one of the originator cells, cancer progresses when the mutant cell lines continue to gain more and more mutations, and finally, become malignant. For example, chronic myelogenous leukemia (CML) develops initially as a non-lethal increase in white blood cells, which progressively...
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Distinctive Features of Adult Stem Cells vs Cancer Stem Cells01:18

Distinctive Features of Adult Stem Cells vs Cancer Stem Cells

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A stem cell is an unspecialized cell that can divide without limit as needed and can, under specific conditions, differentiate into specialized cells.
Adult stem cells
Adult stem cells are tissue-specific; hence, they divide to develop the tissue from which they originate. One type of adult stem cell is the epithelial stem cell, which gives rise to the keratinocytes in the multiple layers of epithelial cells in the epidermis of the skin. Adult bone marrow has three distinct types of stem cells:...
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Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
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Generation of CAR-T Cells for Cancer Immunotherapy.

Qumiao Xu1, Hizkia Harto1, Robert Berahovich1

  • 1Promab Biotechnologies, Richmond, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 23, 2018
PubMed
Summary

Chimeric antigen receptor (CAR)-T cell therapy shows promise for cancer treatment. This study details optimized protocols for generating high-quality CAR-T cells for preclinical research and evaluating their effectiveness.

Keywords:
Adoptive cell therapyCancer immunotherapyChimeric antigen receptorLentiviral gene transferT cells

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Area of Science:

  • Immunology
  • Oncology
  • Gene Therapy

Background:

  • Chimeric antigen receptor (CAR)-T cell therapy is a rapidly advancing field in cancer immunotherapy.
  • CAR-T cell therapies have shown significant success in treating B-cell malignancies, leading to regulatory approvals.
  • The efficacy of CAR-T cell therapy relies heavily on the quality of the manufactured cells.

Purpose of the Study:

  • To provide optimized protocols for the generation of CAR-T cells for preclinical studies.
  • To detail methods for lentiviral gene transfer, cell expansion, and CAR expression detection.
  • To outline procedures for evaluating CAR-T cell cytotoxicity in vitro.

Main Methods:

  • Lentiviral gene transfer was employed for T cell engineering.
  • Optimized protocols for CAR-T cell expansion in culture were established.
  • CAR expression was detected, and in vitro cellular cytotoxicity was evaluated.

Main Results:

  • Standardized protocols for CAR-T cell generation were successfully developed.
  • Methods for assessing CAR expression and cytotoxic function were validated.
  • The protocols facilitate the production of CAR-T cells suitable for preclinical evaluation.

Conclusions:

  • Optimized protocols are crucial for the consistent and effective production of CAR-T cells.
  • These detailed methods support preclinical research and development of CAR-T cell therapies.
  • The described approach aids in advancing the quality control of CAR-T cells for cancer immunotherapy.