Metanephric development in serum-free organ culture

In Vitro ◽  
1982 ◽  
Vol 18 (8) ◽  
pp. 675-682 ◽  
Author(s):  
Ellis D. Avner ◽  
Demetrius Ellis ◽  
Thomas Temple ◽  
Ronald Jaffe
Keyword(s):  
Organ Culture ◽  
Serum Free

THE INFLUENCE OF SERUM ON THE UPTAKE, CONVERSION AND ACTION OF DIHYDROTESTOSTERONE IN RAT PROSTATE GLANDS IN ORGAN CULTURE

1975 ◽  
Vol 64 (2) ◽  
pp. 289-297 ◽  
Author(s):  
ILSE LASNITZKI ◽  
HILARY R. FRANKLIN
Keyword(s):  
Organ Culture ◽  
Horse Serum ◽  
Target Tissue ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Human Pregnancy ◽  
Human Male ◽  
Columnar Cells

SUMMARY The influence of serum on the uptake, conversion and action of dihydrotestosterone in relation to the sex steroid binding protein, TeBG, has been investigated in rat ventral prostates in organ culture. The organs were incubated with [1,2-3H]dihydrotestosterone in: (1) serum-free medium, (2) horse serum, foetal and newborn bovine serum or (3) human male and human pregnancy serum. With all sera the uptake of dihydrotestosterone fell with rising serum concentration, at first steeply and then more gradually. At the same concentration, the uptake was significantly lower in explants incubated with human pregnancy serum than in those kept with human male serum. The conversion of dihydrotestosterone to androstanediol followed the same pattern and less androstanediol was formed in the presence of pregnancy serum. Since pregnancy serum contains higher amounts of TeBG than male serum, the lowered uptake suggests that only the free hormone was available to the target organ. Addition of unlabelled dihydrotestosterone resulted in a higher uptake than that measured in explants incubated with the labelled steroid only. The effect of the human sera on uptake and conversion was correlated with the androgenic activity of dihydrotestosterone applied at physiological concentrations and expressed as the percentage of secretory columnar cells present. The degree of maintenance closely corresponded to the uptake of the hormone. In serum-free medium, the number of columnar cells approached the values found in vivo, with male serum their number, though reduced, was still substantial, with pregnancy serum it was extremely low. It is concluded that the amounts of TeBG present in serum regulate the supply of the hormone to the target tissue and thus control its biological action.

scholarly journals Development of a Short-Term Canine Full-Thickness Skin Organ Culture Method under Serum-Free Conditions

2016 ◽  
Vol 11 (2) ◽  
pp. 61-69 ◽  
Author(s):  
Francesca Abramo ◽  
Andrea Pirone ◽  
Carla Lenzi ◽  
Maria Federica della Valle ◽  
Silvia Vidali ◽  
...  
Keyword(s):  
Organ Culture ◽  
Culture Method ◽  
Full Thickness ◽  
Short Term ◽  
Serum Free ◽  
Full Thickness Skin ◽  
Thickness Skin ◽  
Serum Free Conditions ◽  
Skin Organ Culture

scholarly journals IGF-I-induced enhancement of contractile response in organ-cultured aortae from diabetic rats is mediated by sustained thromboxane A2 release from endothelial cells

2005 ◽  
Vol 186 (2) ◽  
pp. 367-376 ◽  
Author(s):  
Tsuneo Kobayashi ◽  
Takayuki Matsumoto ◽  
Katsuo Kamata
Keyword(s):  
Endothelial Cells ◽  
Organ Culture ◽  
Contractile Response ◽  
Diabetic Rats ◽  
Receptor Expression ◽  
Diabetic Rat ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Igf I

We have investigated the mechanisms underlying the changes in vascular contractile responsiveness induced by insulin and IGF-I in established streptozotocin-induced diabetic rats. The contractile response to noradrenaline (NA) in organ-cultured diabetic rat aortae cultured with insulin or IGF-I was significantly greater than the corresponding responses in (a) diabetic rat aortae cultured in serum-free medium and (b) control rat aortae cultured with insulin or IGF-I. In aortae from which the endothelium was removed after organ culture the contractile response to NA was greater in those cultured with insulin or IGF-I than in those cultured in serum-free medium. This was not true of aortae endothelium denuded before organ culture. The IGF-I-induced enhancement was prevented by treatment with indomethacin (cyclo-oxygenase inhibitor), SQ29548 (thromboxane (TX) A2 receptor antagonist) or fregrelate (TXA2 synthase inhibitor). IGF-I-induced production of TXB2, a metabolite of TXA2, was greater in diabetic than in control aortae and was attenuated by endothelium denudation, indomethacin or AG1024 (IGF-I receptor inhibitor). The expression of the protein and mRNA for the IGF-I receptor (as assessed by RT-PCR and immunohistochemistry) was markedly increased within endothelial cells in diabetic aortae but only slightly increased within smooth muscle cells (versus control rat aortae). Thus, the NA-induced contractile response in aortae from diabetic rats was enhanced by both insulin and IGF-I and this enhancement may be mediated by sustained cyclo-oxygenase-dependent TXA2 production from endothelial cells. The observed enhancement of IGF-I receptor expression within endothelial cells may be causally related to the potentiation of vascular contractility and the increase in TXA2 production.

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.

Abstract 312: Mechanisms Underlying Impaired KCl-induced Contractility After Long-term Serum-free Organ-culture of Rat Isolated Mesenteric Artery

Hypertension ◽  
2013 ◽  
Vol 62 (suppl_1) ◽  
Author(s):  
Tomoka Morita ◽  
Muneyoshi Okada ◽  
Hideyuki Yamawaki
Keyword(s):  
Smooth Muscle ◽  
Organ Culture ◽  
Mesenteric Artery ◽  
Cell Function ◽  
Mesenteric Arteries ◽  
Muscle Contractility ◽  
Smooth Muscle Contractility ◽  
Serum Free ◽  
Functional Changes

(Background and aim) Organ culture of blood vessels is a useful technique to investigate the long-term effects of drugs. Organ culture in a serum-free condition is so far the best way to maintain differentiated cell function. However some functional changes may occur from freshly isolated blood vessel (fresh) such as decreased contractility. Mammalian/mechanical target of rapamycin (mTOR) complex 1 is atypical serine/threonine kinase which integrates various signals induced by growth factors, stress, energy status, oxygen, and amino acids. Here, we investigated the mechanism of decrease in smooth muscle contractility after long-term serum-free organ culture specifically focusing on mTOR. (Methods and results) Rat isolated mesenteric arteries were cultured for 5 days without (0% serum) or with rapamycin (Rap). In 0% serum, absolute contraction by KCl significantly decreased from fresh (n=21 for fresh, n=7 for 0% serum, p<0.01 ). In Rap, the decreased contraction was significantly normalized (n=7, p<0.05 vs. 0% serum). In both 0% serum and Rap, sensitivity to KCl significantly increased from fresh ( p<0.01 vs. fresh). In 0% serum, mTOR expression significantly increased from fresh (n=8, p<0.01 ) which was significantly normalized by rapamycin (n=8, p<0.01 ). Morphological examinations showed degenerative changes in smooth muscle layer of 0% serum which was improved by rapamycin. In 0% serum, expression of myocardin, a master regulator of smooth muscle gene expression significantly decreased from fresh (n=9, p<0.01 ), which was significantly normalized by rapamycin (n=9, p<0.01 ). (Conclusion) Addition of rapamycin prevented the decreased contractility in serum-free organ cultured mesenteric artery perhaps by normalizing the mTOR and downstream myocardin expression as well as arterial degenerative morphological damage. Further studies are needed to clarify how mTOR controls myocardin expression and also how decreased myocardin expression leads to the decreased smooth muscle contractility. This experimental system may be useful for the hypertension research.

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