Epithelial and mesenchymal cell interactions with extracellular matrix material in vitro

Development ◽  
1969 ◽  
Vol 22 (3) ◽  
pp. 395-405
Author(s):  
H. C. Slavkin ◽  
P. Bringas ◽  
J. Cameron ◽  
R. LeBaron ◽  
L. A. Bavetta
Keyword(s):  
Matrix Material ◽  
Chorioallantoic Membrane ◽  
Tooth Germ ◽  
Mesenchymal Cell ◽  
Basic Rule ◽  
Millipore Filter ◽  
Chick Chorioallantoic Membrane ◽  
Mammalian Embryos ◽  
Tooth Germs

Epidermal organogenesis (thyroid gland, salivary gland, feather, hair, skin, thymus gland, tooth, etc.) generally follows a basic rule; epithelium exhibits well-documented interdependence with adjacent mesenchyme for a specific path of development (Grobstein, 1967, for review). Koch (1967) demonstrated in rodent embryos that isolates of incisor epithelial and mesenchymal tissue, separated by a millipore filter, continued to develop. When homotypic tissues were placed in juxtaposition to the filter, no evidence of continued differentiation was observed. Isolated cervical loop tissues of tooth germs from mammalian embryos have been shown to develop into an entire tooth in vitro (Slavkin & Bavetta, 1968 a; Kollar & Baird, 1969). Our laboratory recently reported that isolated tissue preparations (Slavkin & Bavetta, 1968 a) or cell suspensions (Slavkin, Beierle & Bavetta, 1968) of epithelial and mesenchymal cells from the embryonic cervical loop, in recombination on the chick chorioallantoic membrane (CAM), reconstituted and developed into a tooth germ.

Inhibition of tooth germ differentiation in vitro by diazo-oxo-norleucine (DON)

Development ◽  
1979 ◽  
Vol 50 (1) ◽  
pp. 99-109
Author(s):  
Kirsti Hurmerinta ◽  
Irma Thesleff ◽  
Lauri Saxén
Keyword(s):  
Close Association ◽  
Tooth Germ ◽  
Mesenchymal Cell ◽  
Mouse Embryos ◽  
Control Medium ◽  
Glycoprotein Synthesis ◽  
Odontoblast Differentiation ◽  
Cervical Cells ◽  
Tooth Germs

Molar tooth germs from mouse embryos were studied in a Trowell-type organ culture. After 5 days of culture the odontoblasts had secreted predentine and the ameloblasts had differentiated. When cultured in the presence of 10–50 μm diazo-oxo-norleucine (DON), which is a glutamine analogue, the differentiation of odontoblasts was inhibited, but the teeth looked otherwise healthy. When DON was added after 2 days of culture in control medium (at this tims the odontoblasts in the cuspal area were already differentiated), it did not inhibit predentine secretion, ameloblast differentiation, nor enamel secretion. However, this was seen only in the cuspal area and the boundary to the undifferentiated, more cervical cells was distinct. The results support the concept that the mechanism of the differentiation of odontoblasts is different from that of the ameloblasts. We have shown earlier that a close association between the basement msmbrane and the mesenchymal cells is required for odontoblast differentiation. Because DON inteiferes with glycosaminoglycan and glycoprotein synthesis we suggest that DON inhibits odontoblast differentiation by affecting the mesenchymal cell surface and/or the basement membrane.

scholarly journals The growth in vitro of vaccinia virus in chick embryo chorio-allantoic memberane, minced embryo and cell suspensions

1957 ◽  
Vol 55 (3) ◽  
pp. 347-360 ◽  
Author(s):  
H. B. Maitland ◽  
D. I. Magrath
Keyword(s):  
Chick Embryo ◽  
Vaccinia Virus ◽  
Growth Curve ◽  
Chorioallantoic Membrane ◽  
Growth Cycle ◽  
Cell Suspensions ◽  
Rabbit Skin ◽  
Chick Chorioallantoic Membrane ◽  
Considerable Doubt

The growth curve of rabbit skin-adapted vaccinia virus in the chick chorioallantoic membrane incubated in Hanks' solution showed a drop in titre of virus for about 10 hr. followed by growth. At least 25% of virus, sometimes more, remained infective. A similar fall in titre was observed in heated membranes in which the virus did not grow and this occurred also when membranes, either normal or heated, were infected and disintegrated before incubation.The growth curve of virus in minced chick-embryo was similar to that in chorioallantoic membrane.Virus in cell suspensions prepared from chick embryo and incubated in a nutrient medium showed only a small loss of infectivity before growth in some experiments and rarely dropped below 65–70 % of the original titre in others.These results throw considerable doubt on the view that loss of infectivity preceding growth of vaccinia virus should be interpreted as an essential part of a growth cycle.

scholarly journals A comparative study of the development in vivo and in vitro of rat and rabbit molars

1938 ◽  
Vol 126 (844) ◽  
pp. 315-330 ◽  
Keyword(s):  
Tooth Germ ◽  
The Body ◽  
Extrinsic Factors ◽  
Intrinsic Factors ◽  
Mammalian Teeth ◽  
Tooth Germs ◽  
Developmental Mechanics ◽  
General Influence

Although the normal embryology of mammalian teeth has been carefully studied, little is known of the developmental mechanics of teeth. The present communication is concerned with the problem of cusp formation. The main object of the investigation was to find how far the formation of molar cusps was due to extrinsic factors in the jaw and how far to intrinsic factors in the tooth germ itself. Previous work (Glasstone 1936) had shown that embryonic teeth grown in vitro and removed from the general influence of the body continue to develop. In these earlier experiments the rudiments were explanted when cusps had already appeared but before odontoblasts and dentine had differentiated. In the present experiments the tooth germs were explanted at an earlier stage before the cusps had begun to form, to see whether cusps would develop in vitro in the isolated rudiment and if so whether they would correspond in number, shape and arrangement with those of the normal embryonic tooth.

scholarly journals A novel method of screening combinations of angiostatics identifies bevacizumab and temsirolimus as synergistic inhibitors of glioma-induced angiogenesis

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252233
Author(s):  
Michael I. Dorrell ◽  
Heidi R. Kast-Woelbern ◽  
Ryan T. Botts ◽  
Stephen A. Bravo ◽  
Jacob R. Tremblay ◽  
...  
Keyword(s):  
Side Effects ◽  
Tumor Angiogenesis ◽  
Clinical Success ◽  
Low Cost ◽  
Chorioallantoic Membrane ◽  
Cellular Interactions ◽  
Compensatory Mechanisms ◽  
Chick Chorioallantoic Membrane

Tumor angiogenesis is critical for the growth and progression of cancer. As such, angiostasis is a treatment modality for cancer with potential utility for multiple types of cancer and fewer side effects. However, clinical success of angiostatic monotherapies has been moderate, at best, causing angiostatic treatments to lose their early luster. Previous studies demonstrated compensatory mechanisms that drive tumor vascularization despite the use of angiostatic monotherapies, as well as the potential for combination angiostatic therapies to overcome these compensatory mechanisms. We screened clinically approved angiostatics to identify specific combinations that confer potent inhibition of tumor-induced angiogenesis. We used a novel modification of the ex ovo chick chorioallantoic membrane (CAM) model that combined confocal and automated analyses to quantify tumor angiogenesis induced by glioblastoma tumor onplants. This model is advantageous due to its low cost and moderate throughput capabilities, while maintaining complex in vivo cellular interactions that are difficult to replicate in vitro. After screening multiple combinations, we determined that glioblastoma-induced angiogenesis was significantly reduced using a combination of bevacizumab (Avastin®) and temsirolimus (Torisel®) at doses below those where neither monotherapy demonstrated activity. These preliminary results were verified extensively, with this combination therapy effective even at concentrations further reduced 10-fold with a CI value of 2.42E-5, demonstrating high levels of synergy. Thus, combining bevacizumab and temsirolimus has great potential to increase the efficacy of angiostatic therapy and lower required dosing for improved clinical success and reduced side effects in glioblastoma patients.

scholarly journals Mobilization of shell calcium by the chick chorioallantoic membrane in vitro.

1994 ◽  
Vol 190 (1) ◽  
pp. 141-153
Author(s):  
M J Packard
Keyword(s):  
Domestic Fowl ◽  
Culture Medium ◽  
Chorioallantoic Membrane ◽  
The Other ◽  
Chick Chorioallantoic Membrane ◽  
Petri Dishes ◽  
Specific Degradation ◽  
Donor Eggs

Two explants of shell were removed from each of several fertile eggs of domestic fowl at different times during incubation. The chorioallantoic membrane (CAM) was removed from one of the explants (SHELL ONLY) and was left in situ on the other (SHELL+CAM). Explants were cultured for 24, 48 or 96 h at 37 degrees C and 5% CO2 in air in individual Petri dishes containing Dulbecco's modified Eagle's medium, bovine serum albumin, penicillin and streptomycin. Both SHELL+CAM and SHELL ONLY explants released calcium into the culture medium, but the former released considerably more calcium than the latter. More calcium was released by SHELL+CAM explants taken from older eggs than from younger ones, but the age of the donor eggs did not affect release of calcium by SHELL ONLY explants. In addition, release of calcium by SHELL+CAM explants exceeded that shown by SHELL ONLY explants for multiple 24 h intervals. However, the capacity for sustained release of calcium by SHELL+CAM explants declined with age and maturity of the CAM. Manipulations that lead to the death of the CAM abolish the capacity for SHELL+CAM explants to release more calcium than SHELL ONLY explants. Differential release of calcium by SHELL+CAM explants was not attributable to calcium present in the CAM at the onset of culture or to non-specific degradation of the shell by intracellular constituents released as a result of the death of the CAM. Taken in concert, these results indicate that the CAM mobilizes calcium from the eggshell during in vitro culture.

scholarly journals Angiogenic property of silk fibroin scaffolds with adipose-derived stem cells on chick chorioallantoic membrane

2021 ◽  
Vol 8 (3) ◽  
Author(s):  
Tanapong Watchararot ◽  
Weerapong Prasongchean ◽  
Peerapat Thongnuek
Keyword(s):  
Stem Cells ◽  
Silk Fibroin ◽  
Pore Diameter ◽  
Chorioallantoic Membrane ◽  
Control Group ◽  
Adipose Derived Stem Cells ◽  
Chick Chorioallantoic Membrane ◽  
Human Adipose Stem Cells

Angiogenesis is a crucial step in tissue regeneration and repair. Biomaterials that allow or promote angiogenesis are thus beneficial. In this study, angiogenic properties of salt-leached silk fibroin (SF) scaffolds seeded with human adipose stem cells (hADSCs) were studied using chick chorioallantoic membrane (CAM) as a model. The hADSC-seeded SF scaffolds (SF-hADSC) with the porosity of 77.34 ± 6.96% and the pore diameter of 513.95 ± 4.99 µm were implanted on the CAM of chick embryos that were on an embryonic day 8 (E8) of development. The SF-hADSC scaffolds induced a spoke-wheel pattern of capillary network indicative of angiogenesis, which was evident since E11. Moreover, the ingrowth of blood vessels into the scaffolds was seen in histological sections. The unseeded scaffolds induced the same extent of angiogenesis later on E14. By contrast, the control group could not induce the same extent of angiogenesis. In vitro cytotoxicity tests and in vivo angioirritative study reaffirmed the biocompatibility of the scaffolds. This work highlighted that the biocompatible SF-hADSC scaffolds accelerate angiogenesis, and hence they can be a promising biomaterial for the regeneration of tissues that require angiogenesis.

Transepithelial calcium transport in the chick chorioallantoic membrane. I. Isolation and characterization of chorionic ectoderm cells

1993 ◽  
Vol 105 (2) ◽  
pp. 369-379 ◽  
Author(s):  
R.E. Akins ◽  
R.S. Tuan
Keyword(s):  
Calcium Binding ◽  
Chorioallantoic Membrane ◽  
Experimental Manipulation ◽  
Transepithelial Transport ◽  
Temperature Sensitive ◽  
Specific Cell ◽  
Chick Chorioallantoic Membrane ◽  
Isolation And Characterization

The chicken eggshell supplies approximately 80% of the calcium found in the hatchling chick. The mobilization of eggshell calcium into the developing embryo involves the transepithelial transport of large amounts of calcium in a development-specific manner. The cells responsible for the transport of eggshell calcium into the embryonic circulation are the ectodermal cells of the chorioallantoic membrane. In this report, we present a method for the isolation and culture of chorioallantoic membrane ectodermal cells, which are amenable to direct experimental manipulation. Cell preparations are characterized with respect to the expression of an ectoderm-specific cell surface marker (transcalcin, a calcium-binding protein), and a specific enzymatic activity (elevated Ca(2+)-activated ATPase). Functional assessment of in vitro cellular calcium uptake by 45Ca2+ tracer kinetics indicates the persistence of a temperature-sensitive, rapid-influx pathway similar to that observed in vivo. The preparations of primary ectodermal cells present an in vitro system applicable to the experimental analysis of calcium metabolism and transport by the chick chorioallantoic membrane.

scholarly journals Biological Effect and Mechanism of the miR-23b-3p/ANXA2 Axis in Pancreatic Ductal Adenocarcinoma

2018 ◽  
Vol 50 (3) ◽  
pp. 823-840 ◽  
Author(s):  
Dan-ming Wei ◽  
Yi-wu Dang ◽  
Zhen-bo Feng ◽  
Lu Liang ◽  
Lu Zhang ◽  
...  
Keyword(s):  
Pancreatic Ductal Adenocarcinoma ◽  
Chorioallantoic Membrane ◽  
Tumor Formation ◽  
Ductal Adenocarcinoma ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Chick Chorioallantoic Membrane ◽  
Pancreatic Cancer Cells

Background/Aims: Accumulating evidence strongly suggests that microRNAs (miRNAs) modulate the expression of known tumor suppressor genes and oncogenes. In the present study, we found that the proliferation and invasion ability of pancreatic ductal adenocarcinoma (PDAC) cells were significantly suppressed by the overexpression of miR-23b-3p. In addition, there are miR-23b-3p binding sites in annexin A2 (ANXA2). Here, we investigated whether miR-23b-3p had an impact on the progression and metastasis of PDAC by targeting ANXA2. Methods: Cell proliferation, migration, and invasion, and cell cycle assays were performed to explore the effect of miR-23b-3p on various malignant phenotypes of pancreatic cancer cells. The size of tumors was observed following miR-23b-3p overexpression in an in vivo chick chorioallantoic membrane assay. Dual-luciferase reporter, quantitative real-time PCR, western blot, and immunohistochemical analyses were used to validate the relationship between miR-23b-3p and ANXA2 in vitro. Results: We observed that miR-23b-3p could bind specifically to the 3′ untranslated region of ANXA2 and inhibit its expression. MiR-23b-3p overexpression downregulated the expression of ANXA2 mRNA in PDAC cells and limited the size of tumors or even prevented tumor formation. In addition, there was a negative correlation between miR-23b-3p expression and ANXA2 protein expression in clinical specimens. Conclusion: MiR-23b-3p inhibits the development and progression of PDAC by regulating ANXA2 directly.

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