Comparison of Surgical and Cadaveric Intestine as a Source of Crypt Culture in Humans.

Autor:	Scott A; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA., Olack B; Department of Diabetes and Cancer Discovery Science, 20220City of Hope, Integrated Islet Distribution Program and Intestinal Stem Cell Consortium, Coordinating Center, Duarte, CA, USA., Rouch JD; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA., Khalil HA; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA., Kokubun BA; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA., Lei NY; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA., Wang J; Division of Gastroenterology and Nutrition, Department of Pediatrics, Mattel Children's Hospital and the David Geffen School of Medicine, 12222UCLA Los Angeles, CA, USA., Solorzano S; Division of Gastroenterology and Nutrition, Department of Pediatrics, Mattel Children's Hospital and the David Geffen School of Medicine, 12222UCLA Los Angeles, CA, USA., Lewis M; Department of Pathology, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, CA, USA., Dunn JCY; Department of Surgery, 6429Stanford University School of Medicine, Stanford, CA, USA., Stelzner MG; Department of Surgery, David Geffen School of Medicine, 12222UCLA, Los Angeles, CA, USA.; Department of Surgery, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, CA, USA., Niland JC; Department of Diabetes and Cancer Discovery Science, 20220City of Hope, Integrated Islet Distribution Program and Intestinal Stem Cell Consortium, Coordinating Center, Duarte, CA, USA., Martín MG; Division of Gastroenterology and Nutrition, Department of Pediatrics, Mattel Children's Hospital and the David Geffen School of Medicine, 12222UCLA Los Angeles, CA, USA.; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, 12222UCLA, Los Angeles, CA, USA.
Jazyk:	angličtina
Zdroj:	Cell transplantation [Cell Transplant] 2020 Jan-Dec; Vol. 29, pp. 963689720903709.
DOI:	10.1177/0963689720903709
Abstrakt:	Human small intestinal crypts are the source of intestinal stem cells (ISCs) that are capable of undergoing self-renewal and differentiation to an epithelial layer. The development of methods to expand the ISCs has provided opportunities to model human intestinal epithelial disorders. Human crypt samples are usually obtained from either endoscopic or discarded surgical samples, and are thereby exposed to warm ischemia, which may impair their in vitro growth as three-dimensional culture as spheroids or enteroids. In this study we compared duodenal samples obtained from discarded surgical samples to those isolated from whole-body preserved cadaveric donors to generate in vitro cultures. We also examined the effect of storage solution (phosphate-buffered saline or University of Wisconsin [UW] solution) as well as multiple storage times on crypt isolation and growth in culture. We found that intestinal crypts were successfully isolated from cadaveric tissue stored for up to 144 h post-procurement and also were able to generate enteroids and spheroids in certain media conditions. Surgical samples stored in UW after procurement were sufficiently viable up to 24 h and also allowed the generation of enteroids and spheroids. We conclude that surgical samples stored for up to 24 h post-procurement in UW solution allowed for delayed crypt isolation and viable in vitro cultures. Furthermore, in situ, hypothermic preservation in cadaveric duodenal samples permitted crypt/ISC isolation, and successful culture of spheroids and enteroids from tissues held for up to 6 days post-procurement.
Databáze:	MEDLINE
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