Isolation of Colonic Crypts That Maintain Structural and Metabolic Viability In Vitro

Gibson, Peter R.; Pol, E van de; Maxwell, Lesley; Gabriel, Anastasia; Doe, William F.

doi:10.1016/0016-5085(89)91549-7

Cited by 71 publications

(39 citation statements)

References 7 publications

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“…Primary cultures of CEC were prepared as described previously by this laboratory with minor modifications (19). Briefly, fresh mucosal biopsies and resected colorectal material were transferred to HBSS without phenol red or glucose.…”

Section: Methodsmentioning

confidence: 99%

Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease.

et al. 1996

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Evidence of in vivo oxidant-induced injury in inflammatory bowel disease (IBD) is largely indirect. Colon epithelial crypt cells (CEC) from paired specimens of histologically normal and inflamed bowel from IBD patients with active disease were examined for altered protein thiol redox status as an indicator of oxidative damage. When CEC preparations from 22 IBD patients were labeled with the reducedthiol-specific probe [14 C]-iodoacetamide (IAM), there was decreased labeling of a number of proteins indicating oxidation of thiol groups in CEC from inflamed mucosa compared to paired normal mucosa, especially the loss of thiol labeling of a 37-kD protein which was almost completely lost. The loss of reduced protein thiol status for the 37-kD band was paralleled by loss of epithelial cell glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) enzyme activity, an enzyme known to contain an essential reduced cysteine (Cys 149 ) at the active site. The identity of the 37-kD protein as GADPH monomer was confirmed by NH 2 -terminal amino acid sequence analysis.To examine whether this type of in vivo injury could be attributed to biologically relevant oxidants produced by inflammatory cells, CEC prepared from normal mucosa were exposed to H 2 O 2 , OCl

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Section: Methodsmentioning

confidence: 99%

Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease.

et al. 1996

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“…The colonic mucosa has proved difficult to study because this tissue is often heavily contaminated by bacteria and fungi and because the proliferative cells constitute a single layer lying deep in the crypts of Leiberkuhn. Despite attempts to develop longterm cultures of colonic epithelial cells from different species (3)(4)(5), proliferation of these cells has not been demonstrated for more than a few days (6,7). There have been attempts to immortalize or transform intestinal epithelial cells by using known immortalizing viral proteins (8).…”

Section: Introductionmentioning

confidence: 99%

Tumorigenic conversion of p53-deficient colon epithelial cells by an activated Ki-ras gene.

Sevignani¹,

Włodarski²,

Kirillova³

et al. 1998

J. Clin. Invest.

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“…Quaroni et al also used collagenase to isolate and culture epithelial cells from small intestine of rat [8] and pig [9]. Gibson et al [48] employed a combination of collagenase and dispase to dissociate colonic mucosa, although the isolated crypts could not be propagated to yield cell lines. One of the major diffi culties encountered in isolating replicative cells from intestine is the presence of mucus and DNA released from damaged nuclei that cause clumping of cells and prevent their attachment.…”

Section: Organ Culturementioning

confidence: 99%

Intestinal Epithelial Cells In Vitro

Chopra

Dombkowski

Stemmer

et al. 2010

Stem Cells and Development

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Recent advances in the biology of stem cells has resulted in signifi cant interest in the development of normal epithelial cell lines from the intestinal mucosa, both to exploit the therapeutic potential of stem cells in tissue regeneration and to develop treatment models of degenerative disorders of the digestive tract. However, the diffi culty of propagating cell lines of normal intestinal epithelium has impeded research into the molecular mechanisms underlying differentiation of stem/progenitor cells into the various intestinal lineages. Several short-term organ/organoid and epithelial cell culture models have been described. There is a dearth of long-term epithelial and/or stem cell cultures of intestine. With an expanding role of stem cells in the treatment of degenerative disorders, there is a critical need for additional efforts to develop in vitro models of stem/progenitor epithelial cells of intestine. The objective of this review is to recapitulate the current status of technologies and knowledge for in vitro propagation of intestinal epithelial cells, markers of the intestinal stem cells, and gene and protein expression profi les of the intestinal cellular differentiation.

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Isolation of Colonic Crypts That Maintain Structural and Metabolic Viability In Vitro

Cited by 71 publications

References 7 publications

Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease.

Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease.

Tumorigenic conversion of p53-deficient colon epithelial cells by an activated Ki-ras gene.

Intestinal Epithelial Cells In Vitro

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