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A Plate-based Cytotoxicity Assay for the Assessment of Rat Placental Natural Killer Cell Cytolytic Function
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Immunology and Infection
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JoVE Journal Immunology and Infection
A Plate-based Cytotoxicity Assay for the Assessment of Rat Placental Natural Killer Cell Cytolytic Function

A Plate-based Cytotoxicity Assay for the Assessment of Rat Placental Natural Killer Cell Cytolytic Function

DOI:
10.3791/58961-v

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10:44 min

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June 02, 2019

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, , ,
1Department of Emergency Medicine,University of Mississippi Medical Center, 2Department of Experimental Therapeutics and Pharmacology,University of Mississippi Medical Center

Chapters

  • 00:04Title
  • 01:28Lymphocyte Cell Isolation from Placentas
  • 02:28Isolation of Natural Killer Cells
  • 06:32Cytotoxicity Assay: Retrieving NK Cells or YAC1 from Culture or Passing Cells
  • 07:51Cytotoxicity Assay: Assay Protocol
  • 09:17Results: Assessment of the Cytotoxicity Assay
  • 09:56Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we provide a detailed methodology to isolate and assess cytotoxic function of natural killer cells from placentas by a colorimetric plate assay. The reduced uterine perfusion pressure rat model of placental ischemia was chosen to demonstrate the antibody-mediated isolation and assessment of the cytotoxic function of natural killer cells.

Tags

Keywords: – Plate-based Cytotoxicity Assay – Rat Placental Natural Killer Cells – Cytolytic Function – Cytotoxic Activity – Inexpensive Method – Specialized Equipment – Hazardous Materials – CD8+ Cytolytic Cells – Target-effector Cell Ratio Optimization – Mononuclear Cell Isolation – Placenta Homogenization – Density Gradient Centrifugation – Biotin-labeled CD3 Antibody – Magnetic Bead Separation

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