Extended analysis of intratumoral heterogeneity of primary osteosarcoma tissue using 3D-in-vivo-tumor-model

2020 ◽  
Vol 76 (2) ◽  
pp. 133-141
Author(s):  
Anna-Lena Feder ◽  
Eric Pion ◽  
Johannes Troebs ◽  
Ulrich Lenze ◽  
Lukas Prantl ◽  
...  
Keyword(s):  
Chorioallantoic Membrane ◽  
Tumor Model ◽  
Genetic Alterations ◽  
Growth Patterns ◽  
Chick Chorioallantoic Membrane ◽  
Primary Osteosarcoma ◽  
Malignant Group ◽  
High Level ◽  
Tumor Profiling

BACKGROUND: Osteosarcomas are a rare, heterogeneous and malignant group of bone tumors that have a high potential for metastasis and aggressive growth patterns. Treatment of metastasized osteosarcoma is often insufficient and research is compromised by problems encountered when culturing cells or analyzing genetic alterations due to the high level of intratumoral and intertumoral heterogeneity. The chick chorioallantoic membrane (CAM) model, a 3D-in-vivo-tumor-model, could potentially facilitate the investigation of osteosarcoma heterogeneity at an individual and highly specified level. OBJECTIVE: Objective was to establish the grafting and transplantation of different primary osteosarcoma tissue parts onto several consecutive CAMs for tumor profiling and investigation of osteosarcoma heterogeneity. METHODS: Various parts of primary osteosarcoma tissue were grafted onto CAMs and were transplanted onto another CAM for five to seven consecutive times, enabling further experimental analyzes. RESULTS: Primary osteosarcoma tissue parts exhibited satisfactory growth patterns and displayed angiogenic development on the CAM. It was possible to graft and transplant different tumor parts several times while the tissue viability was still high and tumor profiling was performed. CONCLUSIONS: Primary osteosarcoma tissue grew on several different CAMs for an extended time period and neovascularization of serial transplanted tumor parts was observed, improving the versatility of the 3D-in-vivo-tumor-model.

scholarly journals Metallothionein-3 promotes cisplatin chemoresistance remodelling in neuroblastoma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Miguel Angel Merlos Rodrigo ◽  
Hana Michalkova ◽  
Vladislav Strmiska ◽  
Berta Casar ◽  
Piero Crespo ◽  
...  
Keyword(s):  
Cisplatin Resistance ◽  
Neuroblastoma Cells ◽  
Chorioallantoic Membrane ◽  
In Vivo Model ◽  
Chick Chorioallantoic Membrane ◽  
High Level ◽  
Cisplatin Chemoresistance ◽  
The Impact ◽  
First Time

AbstractMetallothionein-3 has poorly characterized functions in neuroblastoma. Cisplatin-based chemotherapy is a major regimen to treat neuroblastoma, but its clinical efficacy is limited by chemoresistance. We investigated the impact of human metallothionein-3 (hMT3) up-regulation in neuroblastoma cells and the mechanisms underlying the cisplatin-resistance. We confirmed the cisplatin-metallothionein complex formation using mass spectrometry. Overexpression of hMT3 decreased the sensitivity of neuroblastoma UKF-NB-4 cells to cisplatin. We report, for the first time, cisplatin-sensitive human UKF-NB-4 cells remodelled into cisplatin-resistant cells via high and constitutive hMT3 expression in an in vivo model using chick chorioallantoic membrane assay. Comparative proteomic analysis demonstrated that several biological pathways related to apoptosis, transport, proteasome, and cellular stress were involved in cisplatin-resistance in hMT3 overexpressing UKF-NB-4 cells. Overall, our data confirmed that up-regulation of hMT3 positively correlated with increased cisplatin-chemoresistance in neuroblastoma, and a high level of hMT3 could be one of the causes of frequent tumour relapses.

Assessment of breast cancer primary tumor material in a 3D in vivo model

2021 ◽  
pp. 1-10
Author(s):  
Cynthia Kohl ◽  
Thiha Aung ◽  
Silke Haerteis ◽  
Thomas Papathemelis
Keyword(s):  
Breast Cancer ◽  
Treatment Plan ◽  
Tumor Biology ◽  
Chorioallantoic Membrane ◽  
Tumor Model ◽  
In Vivo Model ◽  
Additional Information ◽  
Chick Chorioallantoic Membrane ◽  
The Individual

BACKGROUND: Breast cancer is the most common malignant tumor in women and highly heterogeneous with a variety of different molecular subtypes. The analysis of the individual tumor biology is necessary to develop a specific and individualized treatment plan for every patient. The chick chorioallantoic membrane (CAM) model, a 3D-in-vivo-tumor-model, could potentially provide a methodology that facilitates the gain of additional information regarding the tumor biology as well as the testing of the tumor’s individual sensitivity to different therapies. OBJECTIVE: The objective was to establish the grafting of different breast cancer primaries onto the CAM for tumor profiling and the investigation of different parameters. METHODS: Breast cancer primary tissue of different patients was grafted onto the CAM. Subsequently, 3D volume and perfusion measurements were performed during the engraftment period. Histological analyses of the tumors were carried out after the engraftment period. RESULTS: The grafting of the breast cancer primaries onto the CAM was successful. The tumors remained partially vital and displayed angiogenic development on the CAM. CONCLUSIONS: Breast cancer primary material can be grafted onto the CAM and we observed visible and measurable changes of perfusion over time.

The chick embryo chorioallantoic membrane model: a research approach for ex vivo and in vivo experiments

2021 ◽  
Vol 28 ◽  
Author(s):  
Ana Isabel Fraguas-Sánchez ◽  
Cristina Martín-Sabroso ◽  
Ana Isabel Torres-Suárez
Keyword(s):  
Animal Models ◽  
Chorioallantoic Membrane ◽  
New Drugs ◽  
Biological Research ◽  
Research Areas ◽  
Chick Chorioallantoic Membrane ◽  
Biodistribution Studies ◽  
Toxicological Studies ◽  
Cam Model

Background: The chick chorioallantoic membrane (CAM) model has attracted a great deal of interest in pharmaceutical and biological research as an alternative or complementary in vivo assay to animal models. Traditionally, CAM assay has been widely used to perform some toxicological studies, specifically to evaluate the skin, ocular and embryo toxicity of new drugs and formulations, and perform angiogenesis studies. Due to the possibility to generate the tumors onto the CAM, this model has also become an excellent strategy to evaluate the metastatic potential of different tumours and test the efficacy of novel anticancer therapies in vivo. Moreover, in the recent years, its use has considerably grown in other research areas, including the evaluation of new anti-infective agents, the development of biodistribution studies and tissue engineering research. Objectives: This manuscript provides a critical overview of the use of CAM model in pharmaceutical and biological research, especially to test the toxicity of new drugs and formulations and the biodistribution and the efficacy of novel anticancer and anti-infective therapies, analyzing its advantages and disadvantages compared to animal models. Conclusion: The chick chorioallantoic membrane model shows great utility in several research areas, such as cancer, toxicology, biodistribution studies and anti-infective therapies. In fact, it has become an intermediate stage between in vitro experiments and animal studies, and, in the case of toxicological studies (skin and ocular toxicity), has even replaced the animal models.

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.

scholarly journals Isolation and characterization of an inhibitor of neovascularization from scapular chondrocytes.

1992 ◽  
Vol 119 (2) ◽  
pp. 475-482 ◽  
Author(s):  
M A Moses ◽  
J Sudhalter ◽  
R Langer
Keyword(s):  
Growth Factor ◽  
Potent Inhibitor ◽  
Chorioallantoic Membrane ◽  
Boyden Chamber ◽  
Chick Chorioallantoic Membrane ◽  
Isolation And Characterization ◽  
Apparent Homogeneity ◽  
Sds Page ◽  
And Migration

An inhibitor of neovascularization from the conditioned media of scapular chondrocytes established and maintained in serum-free culture has been isolated and characterized. To determine whether this chondrocyte-derived inhibitor (ChDI) was capable of inhibiting neovascularization in vivo, this protein was assayed in the chick chorioallantoic membrane assay. ChDI was a potent inhibitor of angiogenesis in vivo (4 micrograms = 87% avascular zones). This inhibitor is also an inhibitor of fibroblast growth factor-stimulated capillary endothelial cell (EC) proliferation and migration, as well as being an inhibitor of mammalian collagenase. ChDI significantly suppressed capillary EC proliferation in a dose-dependent, reversible manner with an IC50 (the inhibitory concentration at which 50% inhibition is achieved) of 2.025 micrograms/ml. Inhibition by ChDI of growth factor-stimulated capillary EC migration was also observed using a modified Boyden chamber assay (IC50 = 255 ng/ml). SDS-PAGE analysis followed by silver staining of ChDI purified to apparent homogeneity revealed a single band having an M(r) of 35,550. Gel elution experiments demonstrated that only protein eluting at this molecular weight was anti-angiogenic. These studies are the first demonstration that chondrocytes in culture can produce a highly enriched, potent inhibitor of neovascularization which also inhibits collagenase.

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 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.

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