Isolation and characterization of human primary enterocytes from small intestine using a novel method.
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| Författare | Priti Chougule Gustaf Herlenius Nidia Maritza Hernandez Pradeep B Patil Bo Xu Suchitra Sumitran-Holgersson |
|---|---|
| Publicerad i | Scandinavian journal of gastroenterology |
| Volym | 47 |
| Nummer/häfte | 11 |
| Sidor | 1334-43 |
| ISSN | 1502-7708 |
| Publiceringsår | 2012 |
| Publicerad vid | Institutionen för kliniska vetenskaper, sektionen för kirurgi och kirurgisk gastroforskning, Avdelningen för kirurgi |
| Sidor | 1334-43 |
| Länkar | dx.doi.org/10.3109/00365521.2012.70... |
| Ämneskategorier | Gastroenterologi |
Sammanfattning
Abstract Cell culture studies of enterocytes are important in many
fields. However, there are difficulties in obtaining cell lines from
adult human intestine, such as microbial contamination of cultures from
the tissue samples, short life span of enterocytes, overgrowth of
mesenchymal cells, etc. Various model used to obtain adult intestinal
cell lines are very complex requiring use of feeder layer or gel
matrices. The aim of this study was to establish a novel method for the
simple and reproducible isolation of human enterocytes. Enterocytes were
isolated from SI samples (n = 5) obtained from cadaveric donors using a
mechanical procedure, and separation with immunomagnetic beads coated
with anti-EpCAM antibodies. Light and electron microscopy, flow
cytometry and immunocytochemistry techniques were used to characterize
the isolated cells. Immunohistochemical staining of normal SB biopsies
confirmed that the cell cultures maintained an in vivo phenotype as
reflected in cytokeratin expression CK18, CK20 and expression of
intestine-specific markers such as sucrase isomaltase and maltase
glucoamylase. Furthermore, the cells strongly expressed TLR-5, 6, 7, 8
and 10 and several molecules such as CD40, CD86, CD44, ICAM-1 and HLA-DR
which are important in triggering cell-mediated immune responses. This
novel technique provides a unique in vitro system to study the biology
of enterocytes in normal conditions as well as to study inflammatory
processes in various small bowel disorders.
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