Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes.

Published

Journal Article

The intestines - which contain the largest number of immune cells of any organ in the body - are constantly exposed to foreign antigens, both microbial and dietary. Given an increasing understanding that these luminal antigens help shape the immune response and that education of immune cells within the intestine is critical for a number of systemic diseases, there has been increased interest in characterizing the intestinal immune system. However, many published protocols are arduous and time-consuming. We present here a simplified protocol for the isolation of lymphocytes from the small-intestinal lamina propria, intraepithelial layer, and Peyer's patches that is rapid, reproducible, and does not require laborious Percoll gradients. Although the protocol focuses on the small intestine, it is also suitable for analysis of the colon. Moreover, we highlight some aspects that may need additional optimization depending on the specific scientific question. This approach results in the isolation of large numbers of viable lymphocytes that can subsequently be used for flow cytometric analysis or alternate means of characterization.

Full Text

Duke Authors

Cited Authors

  • Couter, CJ; Surana, NK

Published Date

  • May 8, 2016

Published In

PubMed ID

  • 27213538

Pubmed Central ID

  • 27213538

Electronic International Standard Serial Number (EISSN)

  • 1940-087X

International Standard Serial Number (ISSN)

  • 1940-087X

Digital Object Identifier (DOI)

  • 10.3791/54114

Language

  • eng