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The CAM Assay as an Alternative In Vivo Model for Drug Testing

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Organotypic Models in Drug Development

Part of the book series: Handbook of Experimental Pharmacology ((HEP,volume 265))

Abstract

In the last decade, the chicken chorioallantoic membrane (CAM) assay has been re-discovered in cancer research to study the molecular mechanisms of anti-cancer drug effects. Literature about the CAM assay as an alternative in vivo cancer xenograft model according to the 3R principles has exploded in the last 3 years. Following a summary of the basic knowledge about the chicken embryo, we compare advantages and disadvantages with the classical mouse xenograft model, exemplify established and innovative imaging techniques that are used in the CAM model, and give examples of its successful utilization for studying major hallmarks of cancer such as angiogenesis, proliferation, invasion, and metastasis.

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Abbreviations

4-MU:

4-methyllumbelliferone

5-FU:

5-fluoruracil

CAM:

Chorioallantoic membrane

Cas:

CRISPR-associated

CRISPR/Cas:

Clustered regularly interspaced short palindromic repeats

CT:

Computer tomography

DAPK1:

Death associated protein kinase

DZNep:

Deazaneplanocin

ECT :

Electrochemotherapy

EMT:

Epithelial to mesenchymal transition

EZH2:

Zeste homolog 2

GFP :

Green fluorescent protein

HET-CAM assay:

Hen’s egg test on the CAM

MGUS:

Monoclonal gammopathies of undermined significance

MM:

Multiple myeloma

MMP:

Matrix metalloproteinases

MR :

Magnetic resonance

MRI:

Magnetic resonance imaging

MTT:

Tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OECD:

Organisation for Economic Co-operation and Development

PDcE:

Patient derived chicken egg tumor

PDGFR:

Platelet derived growth factor receptor

PDX:

Patient derived tumor xenograft

PET:

Position electron tomography

RB:

Retinoblastoma

SAHA:

Suberanilohydroxamic acid

VEGFR-2:

Vascular endothelial growth factor receptor-2

ZEB1:

Zinc finger E-box-binding homeobox 1

References

  • Armstrong PB, Quigley JP, Sidebottom E (1982) Transepithelial invasion and intramesenchymal infiltration of the chick embryo chorioallantois by tumor cell lines. Cancer Res 42:1826–1837

    CAS  PubMed  Google Scholar 

  • Auerbach R, Kubai L, Sidky Y (1976) Angiogenesis induction by tumors, embryonic tissues, and lymphocytes. Cancer Res 36:3435–3440

    CAS  PubMed  Google Scholar 

  • Ausprunk DH, Folkman J (1976) Vascular injury in transplanted tissues. Fine structural changes in tumor, adult, and embryonic blood vessels. Virchows Arch B Cell Pathol 21:31–44

    Google Scholar 

  • Ausprunk DH, Knighton DR, Folkman J (1974) Differentiation of vascular endothelium in the chick chorioallantois: a structural and autoradiographic study. Dev Biol 38:237–248

    Article  CAS  PubMed  Google Scholar 

  • Ausprunk DH, Knighton DR, Folkman J (1975) Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Role of host and preexisting graft blood vessels. Am J Pathol 79:597–618

    CAS  PubMed  PubMed Central  Google Scholar 

  • Balciuniene N, Tamasauskas A, Valanciute A, Deltuva V, Vaitiekaitis G, Gudinaviciene I, Weis J, von Keyserlingk DG (2009) Histology of human glioblastoma transplanted on chicken chorioallantoic membrane. Medicina 45:123–131

    Article  PubMed  Google Scholar 

  • Bobek V, Plachy J, Pinterova D, Kolostova K, Boubelik M, Jiang P, Yang M, Hoffman RM (2004) Development of a green fluorescent protein metastatic-cancer chick-embryo drug-screen model. Clin Exp Metastasis 21:347–352

    Article  CAS  PubMed  Google Scholar 

  • Böhm J, Muenzner JK, Caliskan A, Ndreshkjana B, Erlenbach-Wunsch K, Merkel S, Croner R, Rau TT, Geppert CI, Hartmann A, Roehe AV, Schneider-Stock R (2019) Loss of enhancer of zeste homologue 2 (EZH2) at tumor invasion front is correlated with higher aggressiveness in colorectal cancer cells. J Cancer Res Clin Oncol 145:2227–2240

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Brignole C, Marimpietri D, Pastorino F, Nico B, Di Paolo D, Cioni M, Piccardi F, Cilli M, Pezzolo A, Corrias MV, Pistoia V, Ribatti D, Pagnan G, Ponzoni M (2006) Effect of bortezomib on human neuroblastoma cell growth, apoptosis, and angiogenesis. J Natl Cancer Inst 98:1142–1157

    Article  CAS  PubMed  Google Scholar 

  • Busch M, Grosse-Kreul J, Wirtz JJ, Beier M, Stephan H, Royer-Pokora B, Metz K, Dunker N (2017) Reduction of the tumorigenic potential of human retinoblastoma cell lines by TFF1 overexpression involves p53/caspase signaling and miR-18a regulation. Int J Cancer 141:549–560

    Article  CAS  PubMed  Google Scholar 

  • Busch M, Papior D, Stephan H, Dunker N (2018) Characterization of etoposide- and cisplatin-chemoresistant retinoblastoma cell lines. Oncol Rep 39:160–172

    CAS  PubMed  Google Scholar 

  • Chambers AF, Schmidt EE, MacDonald IC, Morris VL, Groom AC (1992) Early steps in hematogenous metastasis of B16F1 melanoma cells in chick embryos studied by high-resolution intravital videomicroscopy. J Natl Cancer Inst 84:797–803

    Article  CAS  PubMed  Google Scholar 

  • Cho C-F, Ablack A, Leong H-S, Zijlstra A, Lewis J (2011) Evaluation of nanoparticle uptake in tumors in real time using intravital imaging. J Vis Exp 51:e2808

    Google Scholar 

  • Cho CF, Shukla S, Simpson EJ, Steinmetz NF, Luyt LG, Lewis JD (2014) Molecular targeted viral nanoparticles as tools for imaging cancer. Methods Mol Biol 1108:211–230

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Costa M, Danesi R, Agen C, Di Paolo A, Basolo F, Del Bianchi S, Del Tacca M (1994) MCF-10A cells infected with the int-2 oncogene induce angiogenesis in the chick chorioallantoic membrane and in the rat mesentery. Cancer Res 54:9–11

    CAS  PubMed  Google Scholar 

  • Cruz A, Parnot C, Ribatti D, Corvol P, Gasc JM (2001) Endothelin-1, a regulator of angiogenesis in the chick chorioallantoic membrane. J Vasc Res 38:536–545

    Article  CAS  PubMed  Google Scholar 

  • DeBord LC, Pathak RR, Villaneuva M, Liu HC, Harrington DA, Yu W, Lewis MT, Sikora AG (2018) The chick chorioallantoic membrane (CAM) as a versatile patient-derived xenograft (PDX) platform for precision medicine and preclinical research. Am J Cancer Res 8:1642–1660

    CAS  PubMed  PubMed Central  Google Scholar 

  • Di Paolo D, Loi M, Pastorino F, Brignole C, Marimpietri D, Becherini P, Caffa I, Zorzoli A, Longhi R, Gagliani C, Tacchetti C, Corti A, Allen TM, Ponzoni M, Pagnan G (2009) Liposome-mediated therapy of neuroblastoma. Methods Enzymol 465:225–249

    Article  PubMed  CAS  Google Scholar 

  • Dohle DS, Pasa SD, Gustmann S, Laub M, Wissler JH, Jennissen HP, Dunker N (2009) Chick ex ovo culture and ex ovo CAM assay: how it really works. J Vis Exp 33:e1620

    Google Scholar 

  • Fiorentzis M, Viestenz A, Siebolts U, Seitz B, Coupland SE, Heinzelmann J (2019) The potential use of electrochemotherapy in the treatment of uveal melanoma: in vitro results in 3D tumor cultures and in vivo results in a chick embryo model. Cancers 11:1344

    Article  CAS  PubMed Central  Google Scholar 

  • Goerttler K (1962) The “teratological basic experiment” on the incubated chick embryo, its possibilities and limitations. Klin Wochenschr 40:809–812

    Article  CAS  PubMed  Google Scholar 

  • Hagedorn M, Javerzat S, Gilges D, Meyre A, de Lafarge B, Eichmann A, Bikfalvi A (2005) Accessing key steps of human tumor progression in vivo by using an avian embryo model. Proc Natl Acad Sci U S A 102:1643–1648

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ishiwata I, Ishiwata C, Soma M, Ono I, Nakaguchi T, Ishikawa H (1988) Tumor angiogenic activity of gynecologic tumor cell lines on the chorioallantoic membrane. Gynecol Oncol 29:87–93

    Article  CAS  PubMed  Google Scholar 

  • Ismail MS, Torsten U, Dressler C, Diederichs JE, Huske S, Weitzel H, Berlien HP (1999) Photodynamic therapy of malignant ovarian tumours cultivated on CAM. Lasers Med Sci 14:91–96

    Article  CAS  PubMed  Google Scholar 

  • Jankovic BD, Isakovic K, Lukic ML, Vujanovic NL, Petrovic S, Markovic BM (1975) Immunological capacity of the chicken embryo. I. Relationship between the maturation of lymphoid tissues and the occurrence of cell-mediated immunity in the developing chicken embryo. Immunology 29:497–508

    CAS  PubMed  PubMed Central  Google Scholar 

  • Janse EM, Jeurissen SH (1991) Ontogeny and function of two non-lymphoid cell populations in the chicken embryo. Immunobiology 182:472–481

    Article  CAS  PubMed  Google Scholar 

  • Jedelska J, Strehlow B, Bakowsky U, Aigner A, Hobel S, Bette M, Roessler M, Franke N, Teymoortash A, Werner JA, Eivazi B, Mandic R (2013) The chorioallantoic membrane assay is a promising ex vivo model system for the study of vascular anomalies. In Vivo 27:701–705

    PubMed  Google Scholar 

  • Kauffmann P, Troeltzsch M, Brockmeyer P, Bohnenberger H, Heidekruger PI, Manzke M, Canis M, Gaayathiri S, Schliephake H, Prantl L, Aung T (2018) First experience of chick chorioallantoic membrane (CAM) assay in the clinical work flow with oral squamous cell carcinoma patients. Clin Hemorheol Microcirc 70:487–494

    Article  CAS  PubMed  Google Scholar 

  • Khokha R, Zimmer MJ, Wilson SM, Chambers AF (1992) Up-regulation of TIMP-1 expression in B16-F10 melanoma cells suppresses their metastatic ability in chick embryo. Clin Exp Metastasis 10:365–370

    Article  CAS  PubMed  Google Scholar 

  • Kim J, Yu W, Kovalski K, Ossowski L (1998) Requirement for specific proteases in cancer cell intravasation as revealed by a novel semiquantitative PCR-based assay. Cell 94:353–362

    Article  CAS  PubMed  Google Scholar 

  • Klagsbrun M, Knighton D, Folkman J (1976) Tumor angiogenesis activity in cells grown in tissue culture. Cancer Res 36:110–114

    CAS  PubMed  Google Scholar 

  • Knighton D, Ausprunk D, Tapper D, Folkman J (1977) Avascular and vascular phases of tumour growth in the chick embryo. Br J Cancer 35:347–356

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Komatsu A, Matsumoto K, Saito T, Muto M, Tamanoi F (2019) Patient derived chicken egg tumor model (PDcE model): current status and critical issues. Cell 8:440

    Article  CAS  Google Scholar 

  • Koop S, Khokha R, Schmidt EE, MacDonald IC, Morris VL, Chambers AF, Groom AC (1994) Overexpression of metalloproteinase inhibitor in B16F10 cells does not affect extravasation but reduces tumor growth. Cancer Res 54:4791–4797

    CAS  PubMed  Google Scholar 

  • Koop S, Schmidt EE, MacDonald IC, Morris VL, Khokha R, Grattan M, Leone J, Chambers AF, Groom AC (1996) Independence of metastatic ability and extravasation: metastatic ras-transformed and control fibroblasts extravasate equally well. Proc Natl Acad Sci U S A 93:11080–11084

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kunz P, Schenker A, Sahr H, Lehner B, Fellenberg J (2019) Optimization of the chicken chorioallantoic membrane assay as reliable in vivo model for the analysis of osteosarcoma. PLoS One 14:e0215312

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leighton J, Nassauer J, Tchao R (1985) The chick embryo in toxicology: an alternative to the rabbit eye. Food Chem Toxicol 23:293–298

    Article  CAS  PubMed  Google Scholar 

  • Leong HS, Steinmetz NF, Ablack A, Destito G, Zijlstra A, Stuhlmann H, Manchester M, Lewis JD (2010) Intravital imaging of embryonic and tumor neovasculature using viral nanoparticles. Nat Protoc 5:1406–1417

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lindner P, Paul S, Eckstein M, Münzner J, Erlenbach-Wuensch K, Ahmed H, Mahadevan V, Brabletz T, Hartmann A, Vera J, Schneider-Stock R (2020) EMT transcription factor ZEB1 alters the epigenetic landscape of colorectal cancer cells. Cell Death Dis 11:147

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lokman NA, Elder AS, Ricciardelli C, Oehler MK (2012) Chick chorioallantoic membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci 13:9959–9970

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lokman NA, Price ZK, Hawkins EK, Macpherson AM, Oehler MK, Ricciardelli C (2019) 4-Methylumbelliferone inhibits Cancer stem cell activation and overcomes chemoresistance in ovarian cancer. Cancers 11:1187

    Article  CAS  PubMed Central  Google Scholar 

  • Lucarelli E, Sangiorgi L, Benassi S, Donati D, Gobbi GA, Picci P, Vacca A, Ribatti D (1999) Angiogenesis in lipoma: an experimental study in the chick embryo chorioallantoic membrane. Int J Mol Med 4:593–596

    CAS  PubMed  Google Scholar 

  • Maiuthed A, Ninsontia C, Erlenbach-Wuensch K, Ndreshkjana B, Muenzner JK, Caliskan A, Husayn AP, Chaotham C, Hartmann A, Vial Roehe A, Mahadevan V, Chanvorachote P, Schneider-Stock R (2018) Cytoplasmic p21 mediates 5-fluorouracil resistance by inhibiting pro-apoptotic Chk2. Cancers (Basel) 10:373–409

    Google Scholar 

  • Lugassy C, Barnhill RL (2007) Angiotropic melanoma and extravascular migratory metastasis: a review. Adv Anat Pathol 14:195–201

    Article  PubMed  Google Scholar 

  • MacDonald IC, Schmidt EE, Morris VL, Chambers AF, Groom AC (1992) Intravital videomicroscopy of the chorioallantoic microcirculation: a model system for studying metastasis. Microvasc Res 44:185–199

    Article  CAS  PubMed  Google Scholar 

  • Mangieri D, Nico B, Benagiano V, De Giorgis M, Vacca A, Ribatti D (2008) Angiogenic activity of multiple myeloma endothelial cells in vivo in the chick embryo chorioallantoic membrane assay is associated to a down-regulation in the expression of endogenous endostatin. J Cell Mol Med 12:1023–1028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marimpietri D, Brignole C, Nico B, Pastorino F, Pezzolo A, Piccardi F, Cilli M, Di Paolo D, Pagnan G, Longo L, Perri P, Ribatti D, Ponzoni M (2007) Combined therapeutic effects of vinblastine and rapamycin on human neuroblastoma growth, apoptosis, and angiogenesis. Clin Cancer Res 13:3977–3988

    Article  CAS  PubMed  Google Scholar 

  • Marimpietri D, Nico B, Vacca A, Mangieri D, Catarsi P, Ponzoni M, Ribatti D (2005) Synergistic inhibition of human neuroblastoma-related angiogenesis by vinblastine and rapamycin. Oncogene 24:6785–6795

    Article  CAS  PubMed  Google Scholar 

  • Marzullo A, Vacca A, Roncali L, Pollice L, Ribatti D (1998) Angiogenesis in hepatocellular carcinoma: an experimental study in the chick embryo chorioallantoic membrane. Int J Oncol 13:17–21

    CAS  PubMed  Google Scholar 

  • Mira E, Lacalle RA, Gomez-Mouton C, Leonardo E, Manes S (2002) Quantitative determination of tumor cell intravasation in a real-time polymerase chain reaction-based assay. Clin Exp Metastasis 19:313–318

    Article  CAS  PubMed  Google Scholar 

  • Monteiro AC, Muenzner JK, Andrade F, Rius FE, Ostalecki C, Geppert CI, Agaimy A, Hartmann A, Fujita A, Schneider-Stock R, Jasiulionis MG (2019) Gene expression and promoter methylation of angiogenic and lymphangiogenic factors as prognostic markers in melanoma. Mol Oncol 13:1433–1449

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mostafa LK, Jones DB, Wright DH (1980) Mechanism of the induction of angiogenesis by human neoplastic lymphoid tissue: studies on the chorioallantoic membrane (CAM) of the chick embryo. J Pathol 132:191–205

    Article  CAS  PubMed  Google Scholar 

  • Muenzner JK, Kunze P, Lindner P, Polaschek S, Menke K, Eckstein M, Geppert CI, Chanvorachote P, Baeuerle T, Hartmann A, Schneider-Stock R (2018) Generation and characterization of hepatocellular carcinoma cell lines with enhanced cancer stem cell potential. J Cell Mol Med 22:6238–6248

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ndreshkjana B, Capci A, Klein V, Chanvorachote P, Muenzner JK, Huebner K, Steinmann S, Erlenbach-Wuensch K, Geppert CI, Agaimy A, Ballout F, El-Baba C, Gali-Muhtasib H, Roehe AV, Hartmann A, Tsogoeva SB, Schneider-Stock R (2019) Combination of 5-fluorouracil and thymoquinone targets stem cell gene signature in colorectal cancer cells. Cell Death Dis 10:379

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ossowski L (1988a) In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface-bound urokinase. J Cell Biol 107:2437–2445

    Article  CAS  PubMed  Google Scholar 

  • Ossowski L (1988b) Plasminogen activator dependent pathways in the dissemination of human tumor cells in the chick embryo. Cell 52:321–328

    Article  CAS  PubMed  Google Scholar 

  • Ossowski L, Reich E (1980) Experimental model for quantitative study of metastasis. Cancer Res 40:2300–2309

    CAS  PubMed  Google Scholar 

  • Pacini S, Punzi T, Gulisano M, Ruggiero M (2008) Friend erythroleukemia cells induce angiogenesis in chick embryo chorioallantoic membrane and in human umbilical vein endothelial cells. Biol Res 41:109–117

    Article  PubMed  Google Scholar 

  • Palczak R, Splawinski J (1989) Angiogenic activity and neovascularization in adenocarcinoma of endometrium. Int J Gynaecol Obstet 29:343–357

    Article  CAS  PubMed  Google Scholar 

  • Petruzzelli GJ, Snyderman CH, Johnson JT, Myers EN (1993) Angiogenesis induced by head and neck squamous cell carcinoma xenografts in the chick embryo chorioallantoic membrane model. Ann Otol Rhinol Laryngol 102:215–221

    Article  CAS  PubMed  Google Scholar 

  • Ria R, Loverro G, Vacca A, Ribatti D, Cormio G, Roccaro AM, Selvaggi L (2002) Angiogenesis extent and expression of matrix metalloproteinase-2 and -9 agree with progression of ovarian endometriomas. Eur J Clin Investig 32:199–206

    Article  CAS  Google Scholar 

  • Ribatti D (2010) The chick embryo chorioallantoic membrane in the study of angiogenesis and metastasis. Springer, Dordrecht

    Book  Google Scholar 

  • Ribatti D (2014) The chick embryo chorioallantoic membrane as a model for tumor biology. Exp Cell Res 328:314–324

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Alessandri G, Baronio M, Raffaghello L, Cosimo E, Marimpietri D, Montaldo PG, De Falco G, Caruso A, Vacca A, Ponzoni M (2001a) Inhibition of neuroblastoma-induced angiogenesis by fenretinide. Int J Cancer 94:314–321

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, De Falco G, Nico B, Ria R, Crivellato E, Vacca A (2003) In vivo time-course of the angiogenic response induced by multiple myeloma plasma cells in the chick embryo chorioallantoic membrane. J Anat 203:323–328

    Article  PubMed  PubMed Central  Google Scholar 

  • Ribatti D, Gualandris A, Bastaki M, Vacca A, Iurlaro M, Roncali L, Presta M (1997) New model for the study of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane: the gelatin sponge/chorioallantoic membrane assay. J Vasc Res 34:455–463

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Gualandris A, Belleri M, Massardi L, Nico B, Rusnati M, Dell’Era P, Vacca A, Roncali L, Presta M (1999) Alterations of blood vessel development by endothelial cells overexpressing fibroblast growth factor-2. J Pathol 189:590–599

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Nico B, Cimpean AM, Raica M, Crivellato E, Ruggieri S, Vacca A (2013) B16-F10 melanoma cells contribute to the new formation of blood vessels in the chick embryo chorioallantoic membrane through vasculogenic mimicry. Clin Exp Med 13:143–147

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Nico B, Morbidelli L, Donnini S, Ziche M, Vacca A, Roncali L, Presta M (2001b) Cell-mediated delivery of fibroblast growth factor-2 and vascular endothelial growth factor onto the chick chorioallantoic membrane: endothelial fenestration and angiogenesis. J Vasc Res 38:389–397

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Raffaghello L, Pastorino F, Nico B, Brignole C, Vacca A, Ponzoni M (2002) In vivo angiogenic activity of neuroblastoma correlates with MYCN oncogene overexpression. Int J Cancer 102:351–354

    Article  CAS  PubMed  Google Scholar 

  • Ribatti D, Vacca A, Bertossi M, De Benedictis G, Roncali L, Dammacco F (1990) Angiogenesis induced by B-cell non-Hodgkin’s lymphomas. Lack of correlation with tumor malignancy and immunologic phenotype. Anticancer Res 10:401–406

    CAS  PubMed  Google Scholar 

  • Romanoff AL (1960) The avian embryo, structural and functional development. Macmillan, New York

    Google Scholar 

  • Rous P, Murphy JB (1911) Tumor implantations in the developing embryo. J Am Med Assoc 56:741

    Google Scholar 

  • Rovithi M, Avan A, Funel N, Leon LG, Gomez VE, Wurdinger T, Griffioen AW, Verheul HM, Giovannetti E (2017) Development of bioluminescent chick chorioallantoic membrane (CAM) models for primary pancreatic cancer cells: a platform for drug testing. Sci Rep 7:44686

    Article  PubMed  PubMed Central  Google Scholar 

  • Scher C, Haudenschild C, Klagsbrun M (1976) The chick chorioallantoic membrane as a model system for the study of tissue invasion by viral transformed cells. Cell 8:373–382

    Article  CAS  PubMed  Google Scholar 

  • Sheela S, Riccardi VM, Ratner N (1990) Angiogenic and invasive properties of neurofibroma Schwann cells. J Cell Biol 111:645–653

    Article  CAS  PubMed  Google Scholar 

  • Shimada K, Nakamura M, Ishida E, Higuchi T, Tanaka M, Ota I, Konishi N (2007) c-Jun NH2 terminal kinase activation and decreased expression of mitogen-activated protein kinase phosphatase-1 play important roles in invasion and angiogenesis of urothelial carcinomas. Am J Pathol 171:1003–1012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shioda T, Munn LL, Fenner MH, Jain RK, Isselbacher KJ (1997) Early events of metastasis in the microcirculation involve changes in gene expression of cancer cells. Tracking mRNA levels of metastasizing cancer cells in the chick embryo chorioallantoic membrane. Am J Pathol 150:2099–2112

    CAS  PubMed  PubMed Central  Google Scholar 

  • Skowron MA, Sathe A, Romano A, Hoffmann MJ, Schulz WA, van Koeveringe GA, Albers P, Nawroth R, Niegisch G (2017) Applying the chicken embryo chorioallantoic membrane assay to study treatment approaches in urothelial carcinoma. Urol Oncol 35:544.e511–544.e523

    Article  Google Scholar 

  • Steiling W, Bracher M, Courtellemont P, de Silva O (1999) The HET-CAM, a useful in vitro assay for assessing the eye irritation properties of cosmetic formulations and ingredients. Toxicol In Vitro 13:375–384

    Article  CAS  PubMed  Google Scholar 

  • Steinmann S, Kunze P, Hampel C, Eckstein M, Bramsen JB, Muenzner JK, Carle B, Ndreshkjana B, Kemenes S, Gasparini P, Friedrich O, Andersen C, Geppert C, Wang S, Eyuepoglu I, Bäuerle T, Hartmann A, Schneider-Stock R (2019) DAPK1 loss triggers tumor invasion in colorectal tumor cells. Cell Death Dis 10:1–9

    Article  CAS  Google Scholar 

  • Swadi R, Mather G, Pizer BL, Losty PD, See V, Moss D (2018) Optimising the chick chorioallantoic membrane xenograft model of neuroblastoma for drug delivery. BMC Cancer 18:28

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Swadi RR, Sampat K, Herrmann A, Losty PD, See V, Moss DJ (2019) CDK inhibitors reduce cell proliferation and reverse hypoxia-induced metastasis of neuroblastoma tumours in a chick embryo model. Sci Rep 9:9136

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Uloza V, Kuzminiene A, Salomskaite-Davalgiene S, Palubinskiene J, Balnyte I, Uloziene I, Saferis V, Valanciute A (2015) Effect of laryngeal squamous cell carcinoma tissue implantation on the chick embryo chorioallantoic membrane: morphometric measurements and vascularity. Biomed Res Int 2015:629754

    Article  PubMed  PubMed Central  Google Scholar 

  • Vacca A, Ria R, Semeraro F, Merchionne F, Coluccia M, Boccarelli A, Scavelli C, Nico B, Gernone A, Battelli F, Tabilio A, Guidolin D, Petrucci MT, Ribatti D, Dammacco F (2003) Endothelial cells in the bone marrow of patients with multiple myeloma. Blood 102:3340–3348

    Article  CAS  PubMed  Google Scholar 

  • Vacca A, Ribatti D, Iurlaro M, Albini A, Minischetti M, Bussolino F, Pellegrino A, Ria R, Rusnati M, Presta M, Vincenti V, Persico MG, Dammacco F (1998) Human lymphoblastoid cells produce extracellular matrix-degrading enzymes and induce endothelial cell proliferation, migration, morphogenesis, and angiogenesis. Int J Clin Lab Res 28:55–68

    Article  CAS  PubMed  Google Scholar 

  • van der Horst EH, Leupold JH, Schubbert R, Ullrich A, Allgayer H (2004) TaqMan-based quantification of invasive cells in the chick embryo metastasis assay. BioTechniques 37:940–942

    Article  PubMed  Google Scholar 

  • Vu BT, Shahin SA, Croissant J, Fatieiev Y, Matsumoto K, Le-Hoang Doan T, Yik T, Simargi S, Conteras A, Ratliff L, Jimenez CM, Raehm L, Khashab N, Durand JO, Glackin C, Tamanoi F (2018) Chick chorioallantoic membrane assay as an in vivo model to study the effect of nanoparticle-based anticancer drugs in ovarian cancer. Sci Rep 8:8524

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Warnock G, Turtoi A, Blomme A, Bretin F, Bahri MA, Lemaire C, Libert LC, Seret AEJJ, Luxen A, Castronovo V, Plenevaux AREG (2013) In vivo PET/CT in a human glioblastoma chicken chorioallantoic membrane model: a new tool for oncology and radiotracer development. J Nucl Med 54:1782–1788

    Article  CAS  PubMed  Google Scholar 

  • Weber WT, Mausner R (1977) Migration patterns of avian embryonic bone marrow cells and their differentiation to functional T and B cells. Adv Exp Med Biol 88:47–59

    CAS  PubMed  Google Scholar 

  • Wong CH, Siah KW, Lo AW (2019) Estimation of clinical trial success rates and related parameters. Biostatistics 20:273–286

    Article  PubMed  Google Scholar 

  • Zabielska-Koczywas K, Dolka I, Krol M, Zbikowski A, Lewandowski W, Mieczkowski J, Wojcik M, Lechowski R (2017) Doxorubicin conjugated to glutathione stabilized gold nanoparticles (Au-GSH-Dox) as an effective therapeutic agent for feline injection-site sarcomas-chick embryo chorioallantoic membrane study. Molecules 22:253

    Article  PubMed Central  CAS  Google Scholar 

  • Zijlstra A, Mellor R, Panzarella G, Aimes RT, Hooper JD, Marchenko ND, Quigley JP (2002) A quantitative analysis of rate-limiting steps in the metastatic cascade using human-specific real-time polymerase chain reaction. Cancer Res 62:7083–7092

    CAS  PubMed  Google Scholar 

  • Zuo Z, Syrovets T, Genze F, Abaei A, Ma G, Simmet T, Rasche V (2015) High-resolution MRI analysis of breast cancer xenograft on the chick chorioallantoic membrane. NMR Biomed 28:440–447

    Article  PubMed  Google Scholar 

  • Zuo Z, Syrovets T, Wu Y, Hafner S, Vernikouskaya I, Liu W, Ma G, Weil T, Simmet T, Rasche V (2017) The CAM cancer xenograft as a model for initial evaluation of MR labelled compounds. Sci Rep 7:46690

    Article  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Experimental Tumorpathology, Institute of Pathology, University Hospital, Friedrich-Alexander University Erlangen-Nürnberg, Erlangen, Germany

    Regine Schneider-Stock

  2. Department of Basic Medical Sciences, Neurosciences and Sensory Organs, University of Bari Medical School, Bari, Italy

    Domenico Ribatti

Authors
  1. Regine Schneider-Stock
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  2. Domenico Ribatti
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Correspondence to Regine Schneider-Stock .

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Editors and Affiliations

  1. Institut für Pharmazie, FU Berlin, Berlin, Germany

    Monika Schäfer-Korting

  2. School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil

    Silvya Stuchi Maria-Engler

  3. BASF SE, Experimental Toxicology und Ecology, Ludwigshafen am Rhein, Rheinland-Pfalz, Germany

    Robert Landsiedel

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Schneider-Stock, R., Ribatti, D. (2020). The CAM Assay as an Alternative In Vivo Model for Drug Testing. In: Schäfer-Korting, M., Stuchi Maria-Engler, S., Landsiedel, R. (eds) Organotypic Models in Drug Development. Handbook of Experimental Pharmacology, vol 265. Springer, Cham. https://doi.org/10.1007/164_2020_375

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