Blood from patients with CD or controls was collected and PBMCs were isolated using density gradient centrifugation. Lamina propria mononuclear cells (LPMCs) from intestinal biopsies or gut specimen were isolated using the lamina propria kit (Miltenyi Biotec, Bergisch Gladbach, Germany) as previously described.14 (link) CD4+ T cells were isolated with magnetic beads according to the manufacturer’s instructions (Miltenyi Biotec).
Lamina propria kit
Manufactured by Miltenyi Biotec
Sourced in Germany
The Lamina propria kit is a laboratory equipment designed for the isolation and purification of cells from the lamina propria, a layer of connective tissue found in various organs. This kit provides a standardized and efficient method for researchers to extract and analyze cells from this specific tissue microenvironment.
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Lab products found in correlation
5 protocols using lamina propria kit
1
Isolation of CD4+ T cells from PBMCs and Intestinal Biopsies
2
Isolation of PBMCs and LPMCs from UC Patients
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Blood from UC patients [n = 47] or controls [n = 9] was collected and PBMCs were isolated using density gradient centrifugation. LPMCs from intestinal biopsies or gut specimens [n = 83] were isolated using the lamina propria kit [Miltenyi Biotec] as previously described.33 (link) CD4+or CD14+cells were isolated with magnetic beads according to the manufacturer’s instructions [Miltenyi Biotec].
3
Isolation and Purification of Endothelial Cells
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After cleaning faeces, the colon was cut longitudinally and then laterally into pieces of approximately 0.5 cm in length. Single cells were prepared following the Lamina Propria Kit instructions (Miltenyi, 130‐097‐410). Endothelial cells were positively selected using CD31 microbeads (Miltenyi, 130‐097‐418) and LS columns (Miltenyi, 130‐042‐401). Both CD31‐positive and ‐negative fractions were collected and total proteins were quantified using the BCA protein assay (ThermoFisher, 23225).
4
Analyzing Gut Microbiome and Immunity
5
Stomach Digestion with Collagenase
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Stomachs were digested with collagenase As previously described [8] . Briefly, stomachs were cut into ~0.5 mm pieces then centrifuged at 150 g for 5 min. The pellet was resuspended in 7 mL of HBSS containing 0.2% BSA and 0.4 mg/mL collagenase A (Roche) and incubated shaking at 37°C, 120 rpm. Ten mL complete DMEM was added and digested tissue dissociated by three repeats of forceful pipetting using serological pipettes for 5 mins. Alternatively the Lamina Propria Kit and the gentleMACS dissociator (Miltenyi Biotec) was used as per manufacturer's instructions. Cell suspensions were filtered through a 70 μM cell strainer.
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