Epithelial induction of osteogenesis in embryonic chick mandibular mesenchyme studied by transfilter tissue recombinations

Development ◽  
1984 ◽  
Vol 79 (1) ◽  
pp. 225-242
Author(s):  
R. J. van Exan ◽  
B. K. Hall
Keyword(s):  
Epithelial Cell ◽  
Basal Lamina ◽  
Chorioallantoic Membrane ◽  
Embryonic Chick ◽  
In Ovo ◽  
Cell Product ◽  
Tissue Separation ◽  
Inductive Mechanism ◽  
Millipore Filters ◽  
Sem Studies

The initiation of osteogenesis in the mandibular mesenchyme of the embryonic chick at 7 days is dependent upon an epithelial induction which occurs in the mandible up to the fourth day in ovo. In the present study, transfilter tissue recombinations were used to study this inductive mechanism. The epithelial and mesenchymal components of the mandibles were separated before the completion of the induction and recombined to form transfilter explants which were either cultured for 9 days or grafted onto the chorioallantoic membrane for host embryos for 7 days. Control experiments demonstrated that the tissue separation and recombination techniques did not interfere with the normal epithelial induction, and confirmed that mandibular mesenchyme isolated at this stage was incapable of forming bone. Bone was observed in 86 % of the CAM-grafted intact mandible controls and in 80 % of the cultured intact mandible controls. Bone failed to form in the mesenchyme of transfilter explants when Millipore filters with 0·45 μm pores were used. Bone was observed as frequently as in control explants when the mandibular mesenchyme was separated from its epithelium by 0·8 μm or 0·4 μm porosity Nuclepore filters. Only about 30% of the transfilter explants prepared with 0·1 μm porosity Nuclepore filters formed bone and none of the explants prepared with 0·03 μm porosity Nuclepore filters formed bone. SEM studies demonstrated a distinct correlation between the formation of bone in transfilter explants and the ability of the epithelium and mesenchyme to penetrate the pores of the filters. Thus, the present study provides evidence that the site of the induction is restricted to the epithelial—mesenchymal interface, and that the induction is not mediated by a diffusible substance. The nature of the inductive mechanism is discussed with respect to this and other recent studies which suggest that the induction may be mediated by a non-diffusible epithelial cell product resident in the epithelial basal lamina.

Epithelial induction of osteogenesis in embryonic chick mandibular mesenchyme: a possible role for basal lamina

1983 ◽  
Vol 61 (8) ◽  
pp. 967-979 ◽  
Author(s):  
R. J. Van Exan ◽  
B. K. Hall
Keyword(s):  
Basal Lamina ◽  
Mesenchymal Cell ◽  
Embryonic Chick ◽  
Cell Contacts ◽  
Ultrastructural Studies ◽  
Extracellular Products ◽  
Series Of Experiments ◽  
Inductive Mechanism

The initiation of osteogenesis at 7 days in the embryonic chick mandibular mesenchyme depends on an epithelial induction in the mandible to day 4. This article reviews a series of experiments conducted to study the nature of this inductive mechanism. Transfilter tissue recombinations were used to determine whether direct tissue apposition was required for induction. Ultrastructural studies of the epithelial–mesenchymal interface were conducted to see if direct epithelial–mesenchymal cell–cell contacts occurred during the inductive stage in vivo. Epithelial cells were cultured on Millipore filters for 28 days and allowed to deposit extracellular products. These products were tested for inductive activity. Findings from these three sets of experiments were discussed with respect to the inductive mechanism. Our results indicate that the induction is not mediated by a diffusible substance and that direct apposition of the two tissues is required. The mechanism of induction, however, does not require direct epithelial–mesenchymal cell to cell contacts. This suggests that a nondiffusible component of the extracellular matrix may be involved. Epithelial extracellular products are inductively active and have the appearance of basal lamina. The active component of the extracellular product is proteinaceous, perhaps collagen, and appears to be situated in the epithelial basal lamina. The role of basal lamina in epithelial–mesenchymal interactions is discussed.

Induction of bone by epithelial cell products

Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 37-46
Author(s):  
Brian K. Hall ◽  
R. J. van Exan
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Epithelial Cell ◽  
Basal Lamina ◽  
Inductive Interaction ◽  
Extracellular Products ◽  
Epithelial Cultures ◽  
Vertebrate Embryos ◽  
Induce Bone Formation ◽  
Millipore Filters

The bones of the head and face of vertebrate embryos only form after their progenitor cells have undergonean inductive interaction with embryonic epithelia. We have investigated whether epithelial cell products can substitute for epithelia in allowing mandibular ectomesenchyme to form bone. Mandibular epithelia from embryonic chicks were cultured on Millipore filters for 28 days to allow them to deposit an extracellular matrix, shown by electron microscopy to be a basal lamina-like material. Mandibular ectomesenchymal cells formed bone when placed on to these epithelial extracellular products and grafted to chorioallantoic membranes of host embryos. Treatment of epithelial cultures with trypsin or l-azetidine-carboxylic acid removed both the extracellular products and their ability to induce bone formation. Hyaluronidase treatment did neither. We concluded that a proteinaceous component of epithelial basal lamina provides a sufficient inductive stimulus to initiate differentiation of bone within mandibular ectomesenchyme.

scholarly journals The Role of Pre-Clinical 3-Dimensional Models of Osteosarcoma

2020 ◽  
Vol 21 (15) ◽  
pp. 5499
Author(s):  
Hannah L. Smith ◽  
Stephen A. Beers ◽  
Juliet C. Gray ◽  
Janos M. Kanczler
Keyword(s):  
Three Dimensional ◽  
Chorioallantoic Membrane ◽  
3D Models ◽  
In Ovo ◽  
Future Directions ◽  
3 Dimensional ◽  
In Vivo Animal Models

Treatment for osteosarcoma (OS) has been largely unchanged for several decades, with typical therapies being a mixture of chemotherapy and surgery. Although therapeutic targets and products against cancer are being continually developed, only a limited number have proved therapeutically active in OS. Thus, the understanding of the OS microenvironment and its interactions are becoming more important in developing new therapies. Three-dimensional (3D) models are important tools in increasing our understanding of complex mechanisms and interactions, such as in OS. In this review, in vivo animal models, in vitro 3D models and in ovo chorioallantoic membrane (CAM) models, are evaluated and discussed as to their contribution in understanding the progressive nature of OS, and cancer research. We aim to provide insight and prospective future directions into the potential translation of 3D models in OS.

Nerve-dependent accumulation of myosin light chain 3 in developing limb musculature

Development ◽  
1987 ◽  
Vol 101 (4) ◽  
pp. 673-684
Author(s):  
P.A. Merrifield ◽  
I.R. Konigsberg
Keyword(s):  
Neural Tube ◽  
Light Chain ◽  
Limb Development ◽  
Myosin Light Chain ◽  
Chorioallantoic Membrane ◽  
In Ovo ◽  
Fast Myosin ◽  
Limb Musculature

Myosin alkali light chain accumulation in developing quail limb musculature has been analysed on immunoblots using a monoclonal antibody which recognizes an epitope common to fast myosin light chain 1 (MLC1f) and fast myosin light chain 3 (MLC3f). The limb muscle of early embryos (i.e. up to day 10 in ovo) has a MLC profile similar to that observed in myotubes cultured in vitro; although MLC1f is abundant, MLC3f cannot be detected. MLC3f is first detected in 11-day embryos. To determine whether this alteration in MLC3f accumulation is nerve or hormone dependent, limb buds with and without neural tube were cultured as grafts on the chorioallantoic membrane of chick hosts. Although differentiated muscle develops in both aneural and innervated grafts, innervated grafts contain approximately three times as much myosin as aneural grafts. More significantly, although aneural grafts reproducibly accumulate normal levels of MLC1f, they fail to accumulate detectable levels of MLC3f. In contrast, innervated grafts accumulate both MLC1f and MLC3f, suggesting that the presence of neural tube in the graft promotes the maturation, as well as the growth, of muscle tissue. This is the first positive demonstration that innervation is necessary for the accumulation of MLC3f that occurs during normal limb development in vivo.

scholarly journals Secretome of Senescent Adipose-Derived Mesenchymal Stem Cells Negatively Regulates Angiogenesis

2020 ◽  
Vol 21 (5) ◽  
pp. 1802 ◽  
Author(s):  
Andrey Ratushnyy ◽  
Mariia Ezdakova ◽  
Ludmila Buravkova
Keyword(s):  
Replicative Senescence ◽  
Umbilical Vein ◽  
Chorioallantoic Membrane ◽  
Dot Blot ◽  
In Ovo ◽  
Angiogenic Potential ◽  
Associated Proteins ◽  
Migration Capacity ◽  
Umbilical Vein Endothelial Cells

Nowadays, paracrine regulation is considered as a major tool of mesenchymal stem cell (MSC) involvement in tissue repair and renewal in adults. Aging results in alteration of tissue homeostasis including neovascularization. In this study, we examined the influence of replicative senescence on the angiogenic potential of adipose-derived MSCs (ASCs). Angiogenic activity of conditioned medium (CM) from senescent and “young” ASCs was evaluated in chorioallantoic membrane (CAM) assay in ovo using Japanese quail embryos. Also, the formation of capillary-like tubes by human umbilical vein endothelial cells (HUVECs) in 3D basement membrane matrix “Matrigel” and HUVEC migration capacity were analyzed. Multiplex, dot-blot and gene expression analysis were performed to characterize transcription and production of about 100 angiogenesis-associated proteins. The results point to decreased angiogenic potential of senescent ASC secretome in ovo. A number of angiogenesis-associated proteins demonstrated elevation in CM after long-term cultivation. Meanwhile, VEGF (key positive regulator of angiogenesis) did not change transcription level and concentration in CM. Increasing both pro- (FGF-2, uPA, IL-6, IL-8 etc.) and antiangiogenic (IL-4, IP-10, PF4, Activin A, DPPIV etc.) factors was observed. Some proangiogenic genes were downregulated (IGF1, MMP1, TGFB3, PDGFRB, PGF). Senescence-associated secretory phenotype (SASP) modifications after long-term cultivation lead to attenuation of angiogenic potential of ASC.

scholarly journals ELECTRON PROBE ANALYSIS OF THE CALCIUM DISTRIBUTION IN CELLS OF THE EMBRYONIC CHICK CHORIOALLANTOIC MEMBRANE I. A CRITICAL EVALUATION OF TECHNIQUES

1972 ◽  
Vol 20 (6) ◽  
pp. 401-413 ◽  
Author(s):  
JAMES R. COLEMAN ◽  
A. RAYMOND TEREPKA
Keyword(s):  
Electron Probe ◽  
Critical Evaluation ◽  
Chorioallantoic Membrane ◽  
Embryonic Chick ◽  
Total Calcium ◽  
Electron Probe Analysis ◽  
X Ray ◽  
Chick Chorioallantoic Membrane ◽  
Minor Losses

The chorioallantoic membrane of the developing chick embryo is an epithelium that actively transports calcium. The methodology utilized to prepare this soft tissue for calcium localization with the electron probe x-ray microanalyzer is presented in detail. The preparative procedures are evaluated according to general histochemical principles and in relationship specifically to electron probe investigations. It is shown that the method employed in these studies preserves the normal fine structure of the tissue, prevents selective loss of calcium, permits only minor losses of total calcium and appears to maintain the distribution of calcium that existed in vivo. Examples are presented of artifacts that can be induced during tissue sectioning and mounting procedures. Problems of defining electron probe resolution in biologic specimens are discussed, and the critical importance of evaluating x-ray images in association with simultaneously generated sample current images is emphasized.

Pharmacological analysis of sympathetic function in the embryonic chick heart

1977 ◽  
Vol 232 (3) ◽  
pp. R116-R123
Author(s):  
N. G. Culver ◽  
D. A. Fischman
Keyword(s):  
Sympathetic Nerve ◽  
Sympathetic Innervation ◽  
Sympathetic Nerves ◽  
Nerve Function ◽  
Embryonic Chick ◽  
Neuronal Function ◽  
In Ovo ◽  
Sympathetic Function ◽  
Chick Heart ◽  
Embryonic Chick Heart

Sympathetic nerve cells enter the embryonic chick heart on the fifth day in ovo, but it is uncertain when these nerves become functional. Using pharmacological probes known to affect the embryonic circulation, sympathetic nerve function was examined at various stages of development. Exogenous norepinephrine elicited cardioacceleration in the hearts of embryos with intact extraembryonic circulation both before (stage 20-24) and after (stage 28-32) sympathetic innervation of the heart, and this acceleration could be inhibited by propranolol and practolol. In contrast, ganglionic stimulation with 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) elicited cardioacceleration only after stages 27-28 (i.e., after sympathetic innervation), producing a 25-30% increase in heart rate over the predrug levels of 148.7 +/- 1.8 beats/min. DMPP-elicited positive chronotropy was reduced by beta-receptor antagonists, hexamethonium, guanethidine (GuE), and tetrodotoxin. In preparations of the embryonic thorax in which the innervated heart was separated from brain and adrenal influences, DMPP elicited a GuE-sensitive cardioacceleration. It is concluded that during chick embryonic development, no more than a 1-day interval exists between the appearance of sympathetic nerves in the heart and the onset of neuronal function in that organ.

scholarly journals Morphology of Human Glioblastoma Model Cultured in Ovo

2012 ◽  
Vol 56 (2) ◽  
pp. 261-266 ◽  
Author(s):  
Maciej Szmidt ◽  
Kaja Urbańska ◽  
Marta Grodzik ◽  
Piotr Orłowski ◽  
Ewa Sawosz ◽  
...  
Keyword(s):  
Glioblastoma Multiforme ◽  
Growth Inhibition ◽  
Tumour Growth ◽  
Molecular Mechanisms ◽  
Chorioallantoic Membrane ◽  
Structural Features ◽  
Future Research ◽  
In Ovo ◽  
Tumour Growth Inhibition ◽  
Novel Model

Abstract The aim of this study was the morphological characterisation of glioblastoma multiforme tumour grown in ovo. Tumour cells (U-87 MG) were implanted on the chorioallantoic membrane of chicken egg. The structural features of cultured tumours resembled the spontaneous glioblastoma multiforme; however, the differences were also indicated. Our results confirm applicability of in ovo culture in tumour genesis studies. The described novel model may be profoundly helpful for the future research on molecular mechanisms of tumour growth inhibition.

Expression of basal lamina protein mRNAs in the early embryonic chick eye

2002 ◽  
Vol 447 (3) ◽  
pp. 261-273 ◽  
Author(s):  
Sucai Dong ◽  
Jeffry Landfair ◽  
Manimalha Balasubramani ◽  
Mark E. Bier ◽  
Greg Cole ◽  
...  
Keyword(s):  
Basal Lamina ◽  
Embryonic Chick
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