Protocol detail

Isolation and Purification of Lamina Propria Lymphocytes

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Isolation of LPLs was performed as previously (Li et al., 2017 (link)). The small intestine and large intestine were removed, opened longitudinally, and cut into pieces. After vigorous shaking in HBSS containing EDTA, the supernatants containing epithelial cells and IELs was discarded. The remaining intestinal pieces were digested with collagenase D (Worthington) and pelleted. The pellet was resuspended and placed in a Percoll gradient as described above, and after centrifugation, the interface containing the LPLs was collected and prepared for further analysis.

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Wang Y., Yang H., Jia A., Wang Y., Yang Q., Dong Y., Hou Y., Cao Y., Dong L., Bi Y, & Liu G. (2022). Dendritic cell Piezo1 directs the differentiation of TH1 and Treg cells in cancer. eLife, 11, e79957.

Publication 2022

collagenase D

Centrifugation Collagenase Edta Epithelial cells Hbss Intestinal Isolation Large intestine Lpls Percoll Small intestine

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Variable analysis

independent variables

Isolation of LPLs

dependent variables

LPLs collected at the interface after Percoll gradient centrifugation

control variables

HBSS containing EDTA
Collagenase D (Worthington) for digestion
Percoll gradient centrifugation

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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