Growth of the Vaginal Epithelium of the Mouse in Tissue Culture

L, Martin

doi:10.1677/joe.0.0180334

Journal of Endocrinology

1959

DOI: 10.1677/joe.0.0180334

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Growth of the Vaginal Epithelium of the Mouse in Tissue Culture

Martin L¹

Abstract: The vagina of the immature mouse has been cultured in biological and completely synthetic media for periods up to 72 hr. Growth and cornification of the epithelium occurred in the absence of oestrogens, and was not prevented by the addition of a number of hormones and vitamins to the medium.The significance of the results is discussed, with respect to the mechanism of action of oestrogens.

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Cited by 13 publications

(1 citation statement)

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“…Some investigators showed that explanted rat vaginae (3, 4) and mouse vaginal epithelial outgrowths (2) failed to cornify in response to estradiol, whereas others have reported that the addition of estrogen to the culture medium evoked cornification of vaginal explants from rats (5-8) and mice (9)(10)(11)(12)(13). Martin (14) described growth and cornification of cultured mouse vaginal epithelium in the absence of estrogen. The possible role of the stroma and the difficulty of accurate quantification of increases in cell number are among the problems with both organ and cell culture methods.…”

mentioning

confidence: 99%

Growth of normal mouse vaginal epithelial cells in and on collagen gels.

Iguchi¹,

Uchima²,

Ostrander³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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Sustained growth in primary culture of vaginal epithelial cells from ovariectomized adult BALB/cCrgl mice embedded within or seeded on collagen gel matrix was achieved in a serum-free medium composed of Ham's F-12 medium/Dulbecco's modified Eagle's medium, 1:1 (vol/vol), supplemented with insulin, bovine serum albumin fraction V, epidermal growth factor, cholera toxin, and transferrin. Three-dimensional growth of vaginal epithelial cells occurred inside the collagen gel matrix. Cell numbers increased 4-to 8-fold in collagen gel and about 4-fold on collagen gel after 9-10 days in culture. (3, 4) and mouse vaginal epithelial outgrowths (2) failed to cornify in response to estradiol, whereas others have reported that the addition of estrogen to the culture medium evoked cornification of vaginal explants from rats (5-8) and mice (9-13). Martin (14) described growth and cornification of cultured mouse vaginal epithelium in the absence of estrogen. The possible role of the stroma and the difficulty of accurate quantification of increases in cell number are among the problems with both organ and cell culture methods. Furthermore, in organ culture, tissues degenerate with time; in cell culture, lack of sustained cell division and decreasing viability occur with time (2). To examine the possible direct effect of estrogen on proliferation and cornification of vaginal epithelium, a system is needed for the in vitro culture of isolated epithelial cells.Recently, a collagen gel culture system was developed for mammary epithelial cells that results in sustained growth of mouse, rat, and human cells in primary culture (15)(16)(17)(18)(19)(20)(21). This in vitro system has been used successfully in our studies for the growth of mouse vaginal epithelial cells, as judged by an increase in cell number, and for the possible hormonal regulation of differentiation of these cells in the resulting cultures. In the present communication, we report on the sustained growth of cultured vaginal epithelial cells in a serum-free medium and on the failure of estrogen to stimulate further growth.MATERIALS AND METHODS Epithelial Cell Isolation. Normal vaginae dissected from 50-to 60-day-old BALB/cCrgl mice 6-7 days after ovariectomy were transversely sectioned and then incubated in Hank's balanced salt solution containing 0.1% collagenase (CLS III, 175-184 units per mg; Worthington) and S mg of bovine serum albumin fraction V per ml for 2 hr at 370C in a shaking water bath. Epithelial sheets manually separated from stroma were minced into small clumps with a razor blade on a Teflon block. Cell clumps were collected by centrifugation at 1,000 x g for 5 min.More than 85% of the cells were found to be viable as determined by the trypan blue exclusion test. Cell number was estimated by mixing 1 vol of cell suspension with 9 vol of 0.02% crystal violet in 0.1 M citric acid and counting stained nuclei in a hemocytometer.Preparation of Collagen Gels. Collagen solution and gels were prepared as originally described (22). Briefly, 1 g of rat ta...

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mentioning

confidence: 99%

Growth of normal mouse vaginal epithelial cells in and on collagen gels.

Iguchi¹,

Uchima²,

Ostrander³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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A Microphysiological System with an Anaerobic Air‐Liquid Interface and Functional Mucus Layer for Coculture of Intestinal Bacteria and Primary Human Colonic Epithelium

Kim,

Allbritton

2024

Adv Materials Inter

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Coculture of intestinal bacteria with primary human intestinal epithelium provides a valuable tool for investigating host‐colon bacterial interactions and for testing and screening therapeutics. However, most current intestinal model systems lack key physiological features of the in vivo colon, such as both a proper oxygen microenvironment and a mucus layer. In this work, a new in vitro colonic microphysiological system is demonstrated with a cell‐derived, functional mucus that closely resembles the in vivo colonic mucosa and apical microenvironment by employing an anaerobic air‐liquid interface culture. The human primary colon epithelial cells in this new in vitro system exhibit high cell viability (>98%) with ≈100 µm thick functional mucus layer on average. Successful coculture of model anaerobic gut bacterial strains Lactobacillus rhamnosus GG and Anaerobutyricum hallii without loss in human cell viability demonstrates that this new model can be an invaluable tool for future studies of the impact of commensal and pathogenic bacteria on the colon.

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Estrogen‐independent growth of mouse vaginal epithelium in organ culture

Tsai

Bern

1991

J. Exp. Zool.

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A serum-free vaginal explant culture system was established to investigate the in vitro effect of estrogen on the growth of mouse vaginal epithelium. Vaginal explants were isolated from 40-day-old, ovariectomized BALB/cCrgl mice and cultured in a basal unsupplemented medium or in basal medium plus various doses of 17 beta-estradiol. Explants were processed for histology at the end of culture periods or were given 4-hour pulses of tritiated thymidine at various times and processed for autoradiography. Vaginal epithelium increased 3- to 5-fold in thickness and 2-fold in the number of epithelial cell layers during 72 hours of culture without estrogen; addition of estrogen did not significantly influence epithelial growth. Keratinization of vaginal epithelium occurred within 48 hours of culture in the absence of estrogen, and again addition of estrogen did not accelerate its appearance. Covering the explants with collagen decreased the estrogen-independent growth of vaginal epithelium. Autoradiography showed that ca. 70-90% of basal epithelial cells entered S phase during the initial 4 hours of culture and that this number declined rapidly after 48 hours to ca. 20%. Addition of 1.8 nM 17 beta-estradiol significantly decreased the labelling index of basal cells at 48 hours, but did not affect the labelling index at 24 and 72 hours. Stromal cells were not labelled at any time. Thus, DNA synthesis, cellular proliferation, and differentiation (keratinization) of vaginal epithelium in organ culture occurred without estrogen and were not stimulated by the addition of estrogen.

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Growth of the Vaginal Epithelium of the Mouse in Tissue Culture

Cited by 13 publications

References 8 publications

Growth of normal mouse vaginal epithelial cells in and on collagen gels.

Growth of normal mouse vaginal epithelial cells in and on collagen gels.

A Microphysiological System with an Anaerobic Air‐Liquid Interface and Functional Mucus Layer for Coculture of Intestinal Bacteria and Primary Human Colonic Epithelium

Estrogen‐independent growth of mouse vaginal epithelium in organ culture

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