Effect of hydrocortisone on the maturation of human foetal kidney explants in serum-free organ culture

1989 ◽  
Vol 67 (2-3) ◽  
pp. 121-127 ◽  
Author(s):  
Lyne Bertrand ◽  
Normand Brière
Keyword(s):  
Organ Culture ◽  
Brush Border ◽  
In Vitro Maturation ◽  
Kidney Development ◽  
Thymidine Incorporation ◽  
Brush Border Enzymes ◽  
Serum Free ◽  
Foetal Kidney ◽  
Developing Kidney

The effect of hydrocortisone on the in vitro maturation of human foetal kidney was investigated. Following legal therapeutic abortions, explants of renal cortex from foetuses aged 13–18 weeks were cultured for 5 days in serum-free Leibovitz's L-15 medium at 37 °C in a mixture of 95% air – 5% CO2, without hormone (controls) or with hydrocortisone at concentrations of 12.5, 25, or 50 ng/mL, which are the levels representative of different gestational periods. During the studied period of culture, the overall architecture of the renal structures was preserved without any evident signs of nephrogenesis induced by hydrocortisone. DNA synthesis was measured by incorporation of [3H]thymidine and was stimulated on day 5 by 80% with the addition of hydrocortisone at 12.5 ng/mL, and by 131% with 50 ng/mL. In autoradiograms, the sites of [3H]thymidine incorporation were the same after hydrocortisone addition, but the number of labelled nuclei was higher in 5-day explants supplemented with hydrocortisone at 50 ng/mL. The activities of some brush border enzymes (leucylnaphthylamidase, maltase, and alkaline phosphatase) were not influenced by hydrocortisone when compared with controls. Trehalase activity was decreased on day 5 with 12.5 and 50 ng/mL. A concentration of 12.5 ng/mL diminished γ-glutamyltransferase activity by 29% on day 5. The incorporation of [3H]leucine into proteins was not influenced by any concentration of the glucocorticoid hormone. This study indicates that hydrocortisone directly influences cell proliferation and certain brush border enzymic activities in human developing kidney maintained in organ culture.Key words: hydrocortisone, organ culture, human foetus, kidney, development.

OM-197-MP-AC adjuvant properties: the in vitro maturation of normal and leukemic dendritic cells in a serum-free culture model

Immunobiology ◽  
2004 ◽  
Vol 209 (1-2) ◽  
pp. 67-77 ◽  
Author(s):  
Julie Véran ◽  
Mohamad Mohty ◽  
Béatrice Gaugler ◽  
Carlo Chiavaroli ◽  
Daniel Olive
Keyword(s):  
Dendritic Cells ◽  
In Vitro Maturation ◽  
Serum Free ◽  
Culture Model

scholarly journals The enterocyte microvillus is a vesicle-generating organelle

2009 ◽  
Vol 185 (7) ◽  
pp. 1285-1298 ◽  
Author(s):  
Russell E. McConnell ◽  
James N. Higginbotham ◽  
David A. Shifrin ◽  
David L. Tabb ◽  
Robert J. Coffey ◽  
...  
Keyword(s):  
Brush Border ◽  
Membrane Surface ◽  
Intestinal Lumen ◽  
Intestinal Alkaline Phosphatase ◽  
Brush Border Enzymes ◽  
Membrane Surface Area ◽  
Catalytically Active ◽  
The Right

For decades, enterocyte brush border microvilli have been viewed as passive cytoskeletal scaffolds that serve to increase apical membrane surface area. However, recent studies revealed that in the in vitro context of isolated brush borders, myosin-1a (myo1a) powers the sliding of microvillar membrane along core actin bundles. This activity also leads to the shedding of small vesicles from microvillar tips, suggesting that microvilli may function as vesicle-generating organelles in vivo. In this study, we present data in support of this hypothesis, showing that enterocyte microvilli release unilamellar vesicles into the intestinal lumen; these vesicles retain the right side out orientation of microvillar membrane, contain catalytically active brush border enzymes, and are specifically enriched in intestinal alkaline phosphatase. Moreover, myo1a knockout mice demonstrate striking perturbations in vesicle production, clearly implicating this motor in the in vivo regulation of this novel activity. In combination, these data show that microvilli function as vesicle-generating organelles, which enable enterocytes to deploy catalytic activities into the intestinal lumen.

Microvessel formation from mouse aorta is stimulated in vitro by secreted VEGF and extracts from metanephroi

2003 ◽  
Vol 284 (6) ◽  
pp. C1625-C1632 ◽  
Author(s):  
Tetsu Akimoto ◽  
Marc R. Hammerman
Keyword(s):  
Growth Factors ◽  
Organ Culture ◽  
Fusion Protein ◽  
Thoracic Aorta ◽  
Serum Free ◽  
Hypoxic Conditions ◽  
Thoracic Level ◽  
Anti Vegf ◽  
Mouse Aorta

We have demonstrated that during culture under 5% O2, the addition of recombinant human VEGF or FGF2 to mouse embryonic aorta explants (thoracic level to lateral vessels supplying the mesonephros and metanephros) stimulates microvessel formation. Here we show that microvessel formation is also stimulated by addition to explants of supernatants obtained from metanephroi grown in serum-free organ culture or of metanephroi extracts. Supernatants and extracts from metanephroi grown under hypoxic conditions are more stimulatory than supernatants/extracts from metanephroi grown in room air. VEGF and FGF2 can be detected by using immunohistochemistry in developing nephrons in the cultured renal anlagen. Metanephroi supernatants contain more VEGF if renal anlagen are grown under hypoxic conditions than if they are grown in room air. Metanephros supernatant-stimulated microvessel formation is completely inhibited by soluble sFlt-1 fusion protein or anti-VEGF antibodies (αVEGF). Extract-stimulated microvessel formation is inhibited by αVEGF or anti-FGF2 antibodies, or both. We conclude that metanephroi produce growth factors including VEGF and FGF that enhance microvessel formation from embryonic thoracic aorta in vitro.

scholarly journals EFFECTS OF HYDROCORTISONE ON HUMAN FOETAL KIDNEY IN SERUM-FREE ORGAN CULTURE

1987 ◽  
Vol 21 (4) ◽  
pp. 209A-209A
Author(s):  
Lyne Bertrand ◽  
Normand Brière
Keyword(s):  
Organ Culture ◽  
Serum Free ◽  
Foetal Kidney

Control of Brush Border Enzymes by Dexamethasone in the Fetal Rat Intestine Cultured In Vitro

1982 ◽  
Vol 1 (2) ◽  
pp. 257-266 ◽  
Author(s):  
P. M. Simon-Assmann ◽  
M. Kedinger ◽  
J. F. Grenier ◽  
K. Haffen
Keyword(s):  
Brush Border ◽  
Rat Intestine ◽  
Brush Border Enzymes ◽  
Fetal Rat

scholarly journals Immunohistochemical identification of proliferating cells in organ culture using bromodeoxyuridine and a monoclonal antibody.

1986 ◽  
Vol 34 (6) ◽  
pp. 697-701 ◽  
Author(s):  
G Morstyn ◽  
K Pyke ◽  
J Gardner ◽  
R Ashcroft ◽  
A de Fazio ◽  
...  
Keyword(s):  
Organ Culture ◽  
Cancer Biology ◽  
Thymidine Incorporation ◽  
Optical Data ◽  
Proliferating Cells ◽  
Fetal Mouse ◽  
Mouse Colon ◽  
Dividing Cells ◽  
Immunohistochemical Identification

The ability to measure cell proliferation is important in the study of cancer biology. The usual technique for quantitating proliferating cells in tissue explant and organ culture by detection of [3H]-thymidine incorporation into DNA by autoradiography is tedious and time-consuming. We have developed a technique for identification and quantitation of bromodeoxyuridine (an analogue of thymidine) in cultured tissue explants. Fetal mouse colon explants were exposed in vitro to bromodeoxyuridine (BUdR) or [3H]-thymidine for 3 to 72 hr and then for various periods to unlabeled thymidine. The tissues were stained with a monoclonal anti-bromodeoxyuridine antibody and in parallel [3H]-thymidine incorporation was detected by autoradiography. Incorporation of BUdR was measured by quantitating the amount of pigment deposited over nuclei after immunohistochemical staining, using an optical data digitizer. It was found that both techniques identified proliferating cells. Dividing cells were present both in crypts and in the surrounding stroma in Day 14 fetal mouse colon cultures. The immunohistochemical technique was more rapid and less cumbersome than autoradiography.

Prostatic growth and development are regulated by FGF10

Development ◽  
1999 ◽  
Vol 126 (16) ◽  
pp. 3693-3701
Author(s):  
A.A. Thomson ◽  
G.R. Cunha
Keyword(s):  
Organ Culture ◽  
Seminal Vesicle ◽  
Growth And Development ◽  
Reproductive Tract ◽  
Mesenchymal Cells ◽  
Female Reproductive Tract ◽  
Ventral Prostate ◽  
Serum Free

We have examined the role of Fibroblast Growth Factor 10 (FGF10) during the growth and development of the rat ventral prostate (VP) and seminal vesicle (SV). FGF10 transcripts were abundant at the earliest stages of organ formation and during neonatal organ growth, but were low or absent in growth-quiescent adult organs. In both the VP and SV, FGF10 transcripts were expressed only in a subset of mesenchymal cells and in a pattern consistent with a role as a paracrine epithelial regulator. In the neonatal VP, FGF10 mRNA was expressed initially in mesenchymal cells peripheral to the peri-urethral mesenchyme and distal to the elongating prostatic epithelial buds. At later stages, mesenchymal cells surrounding the epithelial buds also expressed FGF10 transcripts. During induction of the SV, FGF10 mRNA was present in mesenchyme surrounding the lower Wolffian ducts and, at later stages, FGF10 transcripts became restricted to mesenchymal cells subadjacent to the serosa. We investigated whether the FGF10 gene might be regulated by androgens by analysing the levels of FGF10 transcripts in SV and VP organs grown in serum-free organ culture. While FGF10 transcript levels increased after treatment with testosterone in the SV (but not VP), these changes were not sensitive to anti-androgen treatment, and thus it is likely that FGF10 mRNA was not directly regulated by testosterone. Also, FGF10 mRNA was observed in the embryonic female reproductive tract in a position analogous to that of the ventral prostate in males suggesting that FGF10 is not regulated by androgens in vivo. Recombinant FGF10 protein specifically stimulated growth of Dunning epithelial and BPH1 prostatic epithelial cell lines, but had no effect on growth of Dunning stromal cells or primary SV mesenchyme. Furthermore, FGF10 protein stimulated the development of ventral prostate and seminal vesicle organ rudiments in serum-free organ culture. When both FGF10 and testosterone were added to organs in vitro, there was no synergistic induction of development. Additionally, development induced by FGF10 was not inhibited by the addition of the anti-androgen Cyproterone Acetate demonstrating that the effects of FGF10 were not mediated by the androgen receptor. Taken together, our experiments suggest that FGF10 functions as a mesenchymal paracrine regulator of epithelial growth in the prostate and seminal vesicle and that the FGF10 gene is not regulated by androgens

Human foetal kidney explant in serum-free organ culture

1987 ◽  
Vol 176 (1) ◽  
pp. 105-114 ◽  
Author(s):  
Normand Bri�re
Keyword(s):  
Organ Culture ◽  
Serum Free ◽  
Foetal Kidney
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