Successful maintenance of suckling rat ileum in organ culture

Shields, Helen M.; Yedlin, Steven T.; Bair, Frank A.; Goodwin, Carol L.; Alpers, David H.

doi:10.1002/aja.1001550307

Cited by 23 publications

(9 citation statements)

References 27 publications

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“…Furthermore, insulin exerted a sustained effect on crypt cell proliferation [17], It is, however, difficult to conclude whether or not these modifications resulted from a direct action of insulin on the intestinal mucosa or represented a response to a systemic reaction such as for example the transient hypoglycemia caused by insulin [ 19], Because organ culture removes the stud ied tissue from the interacting complexities of the intact organism, it permits a more sys tematic approach to the study of the intrinsic and extrinsic regulators of intestinal epithe lial maturation than is possible in vivo [11,30]. This methodology made it possible to investigate the ontogenetic development of the intestine of the fetal rat, mouse and chick [3,5,6], Few investigations have been de voted to the analysis of intestinal develop-mcnt in vitro using suckling intestine [ 12,28,29]. Since we recently reported the successful maintenance of suckling mouse small intes tine in a serum-free medium [ 16], the present investigation was undertaken to determine the possible direct effect of insulin on the development of the hydrolytic functions of the brush border and on the proliferation of the crypt cells of suckling mouse small intes tine maintained in organ culture.…”

Section: Introductionmentioning

confidence: 99%

Insulin Influences the Maturation and Proliferation of Suckling Mouse Intestinal Mucosa in Serum-Free Organ Culture

Arsenault

Ménard²

1984

Neonatology

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Explants of suckling mouse jejunum have been maintained in serum-free organ culture with or without insulin added to the medium in order to determine the possible direct effect of this hormone on the hydrolytic functions of the brush border and on the proliferation of the crypt cells. The addition of insulin induced the precocious appearance of sucrase activity and increased trehalase, glucoamylase and lactase activities. Alkaline phosphatase activity remained unaffected in the tissue as well as in the medium. An increased DNA content and ³H-thymidine incorporation into DNA were already recorded after 24 h of culture. The mitotic index was significantly increased after 24 h and remained elevated when the culture was extended to 48 h. These results show that insulin directly influences the enzymatic maturation and the proliferation of intestinal epithelial cells of suckling mouse.

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Section: Introductionmentioning

confidence: 99%

Insulin Influences the Maturation and Proliferation of Suckling Mouse Intestinal Mucosa in Serum-Free Organ Culture

Arsenault

Ménard²

1984

Neonatology

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“…As significant changes seem only to be achieved after very long periods, as demonstrated by Eastwood et al (1981), who treated adult rats for 4 weeks, parameters as gland thickness or mucosal weight were not measured in the current study. The edema observed in the mesenchyme in the fetal mucosa may be a consequence from the organ culture system associated with corticosterone effects, as such alteration was not verified when other hormones were added to the medium (Goldfeder and Alvares, 1995), but is similar to that described in other systems (Hayes, 1965;Shields et al, 1979;Finney et al, 1987).…”

Section: Discussionmentioning

confidence: 76%

Cell proliferation and death in the gastric epithelium of developing rats after glucocorticoid treatments

et al. 2000

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Glucocorticoids take part in the intense morphofunctional modifications that occur in the gastric mucosa during fetal and postnatal development. Two studies were designed to evaluate corticoids role in gastric cell proliferation and apoptosis in developing rats: in vivo, using suckling animals; in vitro, using gastric explants obtained from 20-day fetuses. These explants were cultured in DMEM/F12 medium treated or not with 50 ng/ml of corticosterone; after 22 hr, vincristine was added to the medium for 2 hr to block mitosis. The metaphasic index decreased significantly after the 24-hr treatment (controls: 1.52 +/- 0.53; treated: 0.40 +/- 0.21) and apoptotic cells were visualized under light and electron microscopy. Fifteen-day-old rats were treated with hydrocortisone (25 mg/Kg) for 3 days, and injected with BrDU (100 mg/Kg) 1 hr before sacrifice on the 18th day. BrDu-labeled and non-labeled cells were counted to determine the labeling index of epithelial cells. As apoptotic cells are rapidly eliminated, other animals were treated for only 2-3 hr. Sections were investigated for the presence of apoptotic cells, using morphological criteria and TUNEL labeling. Hydrocortisone significantly reduced the labeling index (controls: 15.6 +/- 1.6 vs. treated: 11.7 +/- 1.1), besides altering the body weight gain. Hydrocortisone treatment doubled the number of apoptotic cells after 2 hr, and quadruplicated it after 3 hr. The results demonstrated that glucocorticoids inhibit cell proliferation in the gastric epithelium of fetuses and suckling rats and increase apoptotic rates, suggesting the exit from cell cycle.

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“…Organ culture of the suckling rat ileum was more successful at 25°C than at 37°C (Shields et al 1979). In our system, on the contrary, culture at 37°C was superior to at 24°C, especially under the gas phase of 02 (95%) +CO2 (5%).…”

Section: Discussionmentioning

confidence: 99%

An organ culture method for glandular stomach of new born rat.

Kurokawa¹,

Takahashi²,

Ohno³

et al. 1981

Tohoku J. Exp. Med.

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An organ culture system has been developed for the long-term maintenance of the glandular stomach of new born Wistar rats. The explants from glandular stomach were cultured on Millipore filter in an organ culture dish with various media consisting of sera, buffers and additives under different gas phases of oxygen, carbon dioxide and nitrogen at 37 or 24°C. The culture condition was evaluated by histological examination of explants and was divided into Grade (}}} ), (* ), (+) and (-) depending upon the morphology and growth of the mucosal epithelium. D-MEM plus fetal calf serum (20%) supplemented with hydrocortisone (10,ug/ml) or dexamethasone (5 ug/ml) buffered with sodium bicarbonate and EIepes under a gas phase of 02(95%)+C02(5%) or 02 (45%) +CO2 (5%) +N2 (50%) at 37°C showed the best result. Other supplements tested, such as insulin, ascorbic acid and ferrous sulfate were ineffective. Addition of hydrocortisone or dexamethasone was similarly effective in serum-free media. Isologous rat serum had no advantage over fetal calf serum. Transplantation and autoradiography revealed the viability of the explants up to 35 days in vitro. organ culture; glandular stomach; new born rat Although a number of reports on organ culture of digestive tract organs, such as oral mucosa, esophagus, intestine and colon have been published, those on the stomach have been relatively a few. In spite of our extensive search for references on organ culture of the stomach, we could find only five papers; four on responses of the rat or rabbit gastric mucosa cultured for 24-72 hr to hormonal agents (Sutton and Donaldson 1975;Trier 1976;Yeomans et al. 1976;Lichtenberger et al. 1978), and one on the morphology of human stomach (Hayashi et al. 1975). Similarly, reports on attempts to establish cell lines from the gastric mucosal epithelium have been a few and unsatisfactory (Miller et al. 1973; Kurokawa et al. 1975a, b;Huh, et al. 1977).In this paper, we describe the experiments applied to new born rat glandular stomach in which we have tested various culture media, sera and buffers with several additives under a variety of conditions of gas phase and temperature. The viability of cultured explants was studied by autoradiography and transplantation. The present investigation reports for the first time the long-term organ

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Successful maintenance of suckling rat ileum in organ culture

Cited by 23 publications

References 27 publications

Insulin Influences the Maturation and Proliferation of Suckling Mouse Intestinal Mucosa in Serum-Free Organ Culture

Insulin Influences the Maturation and Proliferation of Suckling Mouse Intestinal Mucosa in Serum-Free Organ Culture

Cell proliferation and death in the gastric epithelium of developing rats after glucocorticoid treatments

An organ culture method for glandular stomach of new born rat.

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