A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery

Sabine Hensler; Claudia Kuehlbach; Bianka Kotkamp; Dario Frey; Margareta Maria Mueller

doi:doi:10.11648/j.bio.20210904.13

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A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery

Sabine Hensler, Claudia Kuehlbach, Bianka Kotkamp, Dario Frey, Margareta Maria Mueller

Published in American Journal of Bioscience and Bioengineering (Volume 9, Issue 4)

Received: 29 June 2021 Accepted: 21 July 2021 Published: 11 August 2021

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Abstract

The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.

Published in	American Journal of Bioscience and Bioengineering (Volume 9, Issue 4)
DOI	10.11648/j.bio.20210904.13
Page(s)	110-122
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Standardized 3D Model, Tumor Stroma Interaction, Inflammatory Cells, Dextran Hydrogel

References

[1]	Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. (2021). Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021 May; 71 (3): 209-249.
[2]	Fennema E, Rivron N, Rouwkema J, van Blitterswijk C, de Boer J. (2013). Spheroid culture as a tool for creating 3D complex tissues. Trends in biotechnology 31 (2): 108-115.
[3]	Van Marion DM, Domanska UM, Timmer-Bosscha H, Walenkamp AM. (2016). Studying cancer metastasis: Existing models, challenges and future perspectives. Critical reviews in oncology/hematology 97: 107-117.
[4]	Sung KE, Beebe DJ. (2014). Microfluidic 3D models of cancer. Advanced drug delivery reviews 79-80: 68-78.
[5]	Van Duinen V, Trietsch SJ, Joore J, Vulto P, Hankemeier T. (2015). Microfluidic 3D cell culture: from tools to tissue models. Current opinion in biotechnology 35: 118-126.
[6]	Mueller MM, Fusenig NE. (2004). Friends or foes - bipolar effects of the tumour stroma in cancer. Nature reviews Cancer 4 (11): 839-849.
[7]	Kuhlbach C, da Luz S, Baganz F, Hass VC, Mueller MM. (2018). A Microfluidic System for the Investigation of Tumor Cell Extravasation. Bioengineering 5 (2).
[8]	Rodrigues T, Kundu B, Silva-Correia J, Kundu SC, Oliveira JM, Reis RL, Correlo VM (2018) Emerging tumor spheroids technologies for 3D in vitro cancer modeling. Pharmacol Ther. 184: 201-211.
[9]	Ravi M, Paramesh V, Kaviya SR, Anuradha E, Solomon FD. (2015). 3D cell culture systems: advantages and applications. Journal of cellular physiology 230 (1): 16-26.
[10]	Edmondson R, Broglie JJ, Adcock AF, Yang L. (2014). Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay and drug development technologies 12 (4): 207-218.
[11]	Sitarski AM, Fairfield H, Falank C, Reagan MR. (2018). 3d Tissue Engineered In Vitro Models Of Cancer In Bone. ACS biomaterials science & engineering 4 (2): 324-336.
[12]	Hughes CS, Postovit LM, Lajoie GA. (2010). Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 10 (9): 1886-1890.
[13]	Vukicevic S, Kleinman HK, Luyten FP, Roberts AB, Roche NS, Reddi AH. (1992). Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Experimental cell research 202 (1): 1-8.
[14]	Augst AD, Kong HJ, Mooney DJ. (2006). Alginate hydrogels as biomaterials. Macromolecular bioscience 6 (8): 623-633.
[15]	Chyzy A, Plonska-Brzezinska ME. (2020) Hydrogel Properties and Their Impact on Regenerative Medicine and Tissue Engineering. Molecules 8; 25 (24): 5795.
[16]	Hirakawa T, Kakunaga T, Fujiki H, Sugimura T. (1982). A new tumor-promoting agent, dihydroteleocidin B, markedly enhances chemically induced malignant cell transformation. Science 216 (4545): 527-529.
[17]	Collins SJ. (1987). The HL-60 promyelocytic leukemia cell line: proliferation, differentiation, and cellular oncogene expression. Blood 70 (5): 1233-1244.
[18]	Fusenig NE, Boukamp P. (1998). Multiple stages and genetic alterations in immortalization, malignant transformation, and tumor progression of human skin keratinocytes. Molecular carcinogenesis 23 (3): 144-158.
[19]	Lederle W, Depner S, Schnur S, Obermueller E, Catone N, Just A, Fusenig NE, Mueller MM. (2011). IL-6 promotes malignant growth of skin SCCs by regulating a network of autocrine and paracrine cytokines. International journal of cancer 128 (12): 2803-2814.
[20]	Depner S, Lederle W, Gutschalk C, Linde N, Zajonz A, Mueller MM. (2014) Cell type specific interleukin-6 induced responses in tumor keratinocytes and stromal fibroblasts are essential for invasive growth. Int J Cancer. 135 (3): 551-62.
[21]	Obermueller E, Vosseler S, Fusenig NE, Mueller MM. (2004). Cooperative autocrine and paracrine functions of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor in the progression of skin carcinoma cells. Cancer research 64 (21): 7801-7812.
[22]	Gutschalk CM, Herold-Mende CC, Fusenig NE, Mueller MM. (2006). Granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor promote malignant growth of cells from head and neck squamous cell carcinomas in vivo. Cancer research 66 (16): 8026-8036.
[23]	Linde N, Gutschalk CM, Hoffmann C, Yilmaz D, Mueller MM. (2012). Integrating macrophages into organotypic co-cultures: a 3D in vitro model to study tumor-associated macrophages. PloS one 7 (7): e40058.
[24]	Brassard-Jollive N, Monnot C, Muller L, Germain S. (2020) In vitro 3D Systems to Model Tumor Angiogenesis and Interactions With Stromal Cells. Front Cell Dev Biol. 2020 Nov 5; 8: 594903.
[25]	Kunz-Schughart LA, Freyer JP, Hofstaedter F, Ebner R. (2004). The use of 3-D cultures for high-throughput screening: the multicellular spheroid model. Journal of biomolecular screening 9 (4): 273-285.
[26]	Fasano MB, Cousart S, Neal S, McCall CE. (1991). Increased expression of the interleukin 1 receptor on blood neutrophils of humans with the sepsis syndrome. The Journal of clinical investigation 88 (5): 1452-1459.
[27]	Ryu JS, Kang JH, Jung SY, Shin MH, Kim JM, Park H, Min DY. (2004). Production of interleukin-8 by human neutrophils stimulated with Trichomonas vaginalis. Infection and immunity 72 (3): 1326-1332.
[28]	Karmakar M, Katsnelson MA, Dubyak GR, Pearlman E. (2016). Neutrophil P2X7 receptors mediate NLRP3 inflammasome-dependent IL-1beta secretion in response to ATP. Nature communications 7: 10555.
[29]	Iula L, Keitelman IA, Sabbione F, Fuentes F, Guzman M, Galletti JG, Gerber PP, Ostrowski M, Geffner JR, Jancic CC and others. (2018). Autophagy Mediates Interleukin-1beta Secretion in Human Neutrophils. Frontiers in immunology 9: 269.
[30]	Fujishima S, Hoffman AR, Vu T, Kim KJ, Zheng H, Daniel D, Kim Y, Wallace EF, Larrick JW, Raffin TA. (1993). Regulation of neutrophil interleukin 8 gene expression and protein secretion by LPS, TNF-alpha, and IL-1 beta. Journal of cellular physiology 154 (3): 478-485.
[31]	Piccard H, Muschel RJ, Opdenakker G. (2012). On the dual roles and polarized phenotypes of neutrophils in tumor development and progression. Critical reviews in oncology/hematology 82 (3): 296-309.
[32]	Jaillon S, Ponzetta A, Di Mitri D, Santoni A, Bonecchi R, Mantovani A. (2020). Neutrophil diversity and plasticity in tumor progression and therapy. Nat Rev Cancer. 20 (9): 485-503.
[33]	Fridlender ZG, Sun J, Kim S, Kapoor V, Cheng G, Ling L, Worthen GS, Albelda SM. (2009). Polarization of tumor-associated neutrophil phenotype by TGF-beta: "N1" versus "N2" TAN. Cancer cell 16 (3): 183-194.
[34]	Jablonska J, Leschner S, Westphal K, Lienenklaus S, Weiss S. (2010). Neutrophils responsive to endogenous IFN-beta regulate tumor angiogenesis and growth in a mouse tumor model. The Journal of clinical investigation 120 (4): 1151-1164.
[35]	Brooks SC, Locke ER, Soule HD. (1973). Estrogen receptor in a human cell line (MCF-7) from breast carcinoma. The Journal of biological chemistry 248 (17): 6251-6253.
[36]	H838 (1996) NCI Navy Medical Oncology Branch Cell Line Supplement J Cell. Bio. Suppl 24.
[37]	Langer R, Tirrell DA. (2004). Designing materials for biology and medicine. Nature 428 (6982): 487-492.
[38]	Meides A, Gutschalk CM, Devel L, Beau F, Czarny B, Hensler S, Neugebauer J, Dive V, Angel P, Mueller MM. (2014). Effects of selective MMP-13 inhibition in squamous cell carcinoma depend on estrogen. International journal of cancer 135 (12): 2749-2759.
[39]	Fridlender ZG, Albelda SM. (2012). Tumor-associated neutrophils: friend or foe? Carcinogenesis 33 (5): 949-955.
[40]	Gregory AD, Houghton AM. (2011). Tumor-associated neutrophils: new targets for cancer therapy. Cancer research 71 (7): 2411-2416.
[41]	Nielsen BS, Timshel S, Kjeldsen L, Sehested M, Pyke C, Borregaard N, Dano K. (1996). 92 kDa type IV collagenase (MMP-9) is expressed in neutrophils and macrophages but not in malignant epithelial cells in human colon cancer. International journal of cancer 65 (1): 57-62.
[42]	Bellocq A, Antoine M, Flahault A, Philippe C, Crestani B, Bernaudin JF, Mayaud C, Milleron B, Baud L, Cadranel J. (1998). Neutrophil alveolitis in bronchioloalveolar carcinoma: induction by tumor-derived interleukin-8 and relation to clinical outcome. The American journal of pathology 152 (1): 83-92.
[43]	Mentzel T, Brown LF, Dvorak HF, Kuhnen C, Stiller KJ, Katenkamp D, Fletcher CD. (2001). The association between tumour progression and vascularity in myxofibrosarcoma and myxoid/round cell liposarcoma. Virchows Archiv: an international journal of pathology 438 (1): 13-22.
[44]	Eck M, Schmausser B, Scheller K, Brandlein S, Muller-Hermelink HK. (2003). Pleiotropic effects of CXC chemokines in gastric carcinoma: differences in CXCL8 and CXCL1 expression between diffuse and intestinal types of gastric carcinoma. Clinical and experimental immunology 134 (3): 508-515.
[45]	Mhawech-Fauceglia P, Kaya G, Sauter G, McKee T, Donze O, Schwaller J, Huard B. (2006). The source of APRIL up-regulation in human solid tumor lesions. Journal of leukocyte biology 80 (4): 697-704.
[46]	Azevedo PO, Paiva AE, Santos GSP, Lousado L, Andreotti JP, Sena IFG, Tagliati CA, Mintz A, Birbrair A. (2018). Cross-talk between lung cancer and bones results in neutrophils that promote tumor progression. Cancer metastasis reviews 37 (4): 779-790.
[47]	Skobe M, Fusenig NE. (1998). Tumorigenic conversion of immortal human keratinocytes through stromal cell activation. Proceedings of the National Academy of Sciences of the United States of America 95 (3): 1050-1055.
[48]	Alemany-Ribes M, Semino CE. (2014). Bioengineering 3D environments for cancer models. Advanced drug delivery reviews 79-80: 40-49.
[49]	Kapalczynska M, Kolenda T, Przybyla W, Zajaczkowska M, Teresiak A, Filas V, Ibbs M, Blizniak R, Luczewski L, Lamperska K. (2018). 2D and 3D cell cultures - a comparison of different types of cancer cell cultures. Archives of medical science: AMS 14 (4): 910-919.
[50]	Levesque SG, Shoichet MS. (2007). Synthesis of enzyme-degradable, peptide-cross-linked dextran hydrogels. Bioconjugate chemistry 18 (3): 874-885.
[51]	Liu Y, Chan-Park MB. (2009). Hydrogel based on interpenetrating polymer networks of dextran and gelatin for vascular tissue engineering. Biomaterials 30 (2): 196-207.
[52]	Fonseca, K. B. Granja, P. L., Barrias, C. C. (2014). Engineering proteolytically-degradable artificial extracellular matrices, Progress in Polymer Science. 39, 2010-2029.
[53]	Yamada KM, Cukierman E. (2007). Modeling tissue morphogenesis and cancer in 3D. Cell 130 (4): 601-610.
[54]	Berning M, Prätzel-Wunder S, Bickenbach JR, Boukamp P. (2015) Three-Dimensional In Vitro Skin and Skin Cancer Models Based on Human Fibroblast-Derived Matrix. Tissue Eng Part C Methods. 21 (9): 958-70.
[55]	Rikken G, Niehues H, van den Bogaard EH. (2020) Organotypic 3D Skin Models: Human Epidermal Equivalent Cultures from Primary Keratinocytes and Immortalized Keratinocyte Cell Lines. Methods Mol Biol. 2154: 45-61.
[56]	Ridky TW, Chow JM, Wong DJ, Khavari PA. (2010). Invasive three-dimensional organotypic neoplasia from multiple normal human epithelia. Nature medicine 16 (12): 1450-1455.
[57]	Benbrook, D. M. (2006). Organotypic cultures represent tumor microenvironment for drug testing, Drug Discovery Today: Disease Models 3, 143–148.
[58]	Bechetoille N, Vachon H, Gaydon A, Boher A, Fontaine T, Schaeffer E, Decossas M, Andre-Frei V, Mueller CG. (2011). A new organotypic model containing dermal-type macrophages. Experimental dermatology 20 (12): 1035-1037.
[59]	Friedl P. (2004). Prespecification and plasticity: shifting mechanisms of cell migration. Current opinion in cell biology 16 (1): 14-23.
[60]	Breslin S, O'Driscoll L. (2016). The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance. Oncotarget 7 (29): 45745-45756.
[61]	Keith B, Simon MC. (2007). Hypoxia-inducible factors, stem cells, and cancer. Cell 129 (3): 465-472.
[62]	Griffith LG, Swartz MA. (2006). Capturing complex 3D tissue physiology in vitro. Nature reviews Molecular cell biology 7 (3): 211-224.
[63]	Nelson CM, Bissell MJ. (2006). Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Annual review of cell and developmental biology 22: 287-309.
[64]	Grinnell F. (2003). Fibroblast biology in three-dimensional collagen matrices. Trends in cell biology 13 (5): 264-269.
[65]	Cukierman E, Pankov R, Yamada KM. (2002). Cell interactions with three-dimensional matrices. Current opinion in cell biology 14 (5): 633-639.
[66]	Amatangelo MD, Bassi DE, Klein-Szanto AJ, Cukierman E. (2005). Stroma-derived three-dimensional matrices are necessary and sufficient to promote desmoplastic differentiation of normal fibroblasts. The American journal of pathology 167 (2): 475-488.
[67]	Thiery JP, Sleeman JP. (2006). Complex networks orchestrate epithelial-mesenchymal transitions. Nature reviews Molecular cell biology 7 (2): 131-142.
[68]	Hay ED. (2005). The mesenchymal cell, its role in the embryo, and the remarkable signaling mechanisms that create it. Developmental dynamics: an official publication of the American Association of Anatomists 233 (3): 706-720.
[69]	Pampaloni F, Reynaud EG, Stelzer EH. (2007). The third dimension bridges the gap between cell culture and live tissue. Nature reviews Molecular cell biology 8 (10): 839-845.
[70]	Bissell MJ, Labarge MA. (2005). Context, tissue plasticity, and cancer: are tumor stem cells also regulated by the microenvironment? Cancer cell 7 (1): 17-23.
[71]	Postovit LM, Costa FF, Bischof JM, Seftor EA, Wen B, Seftor RE, Feinberg AP, Soares MB, Hendrix MJ. (2007). The commonality of plasticity underlying multipotent tumor cells and embryonic stem cells. Journal of cellular biochemistry 101 (4): 908-917.
[72]	Tibbitt MW, Anseth KS. (2009). Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechnology and bioengineering 103 (4): 655-663.
[73]	Braun B, Lange M, Oeckler R, Mueller MM. (2004). Expression of G-CSF and GM-CSF in human meningiomas correlates with increased tumor proliferation and vascularization. Journal of neuro-oncology 68 (2): 131-140.

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APA Style

Sabine Hensler, Claudia Kuehlbach, Bianka Kotkamp, Dario Frey, Margareta Maria Mueller. (2021). A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. American Journal of Bioscience and Bioengineering, 9(4), 110-122. https://doi.org/10.11648/j.bio.20210904.13

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ACS Style

Sabine Hensler; Claudia Kuehlbach; Bianka Kotkamp; Dario Frey; Margareta Maria Mueller. A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. Am. J. BioSci. Bioeng. 2021, 9(4), 110-122. doi: 10.11648/j.bio.20210904.13

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AMA Style

Sabine Hensler, Claudia Kuehlbach, Bianka Kotkamp, Dario Frey, Margareta Maria Mueller. A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. Am J BioSci Bioeng. 2021;9(4):110-122. doi: 10.11648/j.bio.20210904.13

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@article{10.11648/j.bio.20210904.13,
  author = {Sabine Hensler and Claudia Kuehlbach and Bianka Kotkamp and Dario Frey and Margareta Maria Mueller},
  title = {A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery},
  journal = {American Journal of Bioscience and Bioengineering},
  volume = {9},
  number = {4},
  pages = {110-122},
  doi = {10.11648/j.bio.20210904.13},
  url = {https://doi.org/10.11648/j.bio.20210904.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20210904.13},
  abstract = {The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.},
 year = {2021}
}

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TY  - JOUR
T1  - A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery
AU  - Sabine Hensler
AU  - Claudia Kuehlbach
AU  - Bianka Kotkamp
AU  - Dario Frey
AU  - Margareta Maria Mueller
Y1  - 2021/08/11
PY  - 2021
N1  - https://doi.org/10.11648/j.bio.20210904.13
DO  - 10.11648/j.bio.20210904.13
T2  - American Journal of Bioscience and Bioengineering
JF  - American Journal of Bioscience and Bioengineering
JO  - American Journal of Bioscience and Bioengineering
SP  - 110
EP  - 122
PB  - Science Publishing Group
SN  - 2328-5893
UR  - https://doi.org/10.11648/j.bio.20210904.13
AB  - The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.
VL  - 9
IS  - 4
ER  -

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Author Information

Sabine Hensler

Molecular Cell Biology Laboratory and Institute of Technical Medicine, Furtwangen University, Villingen-Schwenningen, Germany
Claudia Kuehlbach

Molecular Cell Biology Laboratory and Institute of Technical Medicine, Furtwangen University, Villingen-Schwenningen, Germany
Bianka Kotkamp

Molecular Cell Biology Laboratory and Institute of Technical Medicine, Furtwangen University, Villingen-Schwenningen, Germany
Dario Frey

Molecular Cell Biology Laboratory and Institute of Technical Medicine, Furtwangen University, Villingen-Schwenningen, Germany
Margareta Maria Mueller

Molecular Cell Biology Laboratory and Institute of Technical Medicine, Furtwangen University, Villingen-Schwenningen, Germany

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APA Style

Sabine Hensler, Claudia Kuehlbach, Bianka Kotkamp, Dario Frey, Margareta Maria Mueller. (2021). A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. American Journal of Bioscience and Bioengineering, 9(4), 110-122. https://doi.org/10.11648/j.bio.20210904.13

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ACS Style

Sabine Hensler; Claudia Kuehlbach; Bianka Kotkamp; Dario Frey; Margareta Maria Mueller. A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. Am. J. BioSci. Bioeng. 2021, 9(4), 110-122. doi: 10.11648/j.bio.20210904.13

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AMA Style

Sabine Hensler, Claudia Kuehlbach, Bianka Kotkamp, Dario Frey, Margareta Maria Mueller. A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery. Am J BioSci Bioeng. 2021;9(4):110-122. doi: 10.11648/j.bio.20210904.13

Copy | Download

@article{10.11648/j.bio.20210904.13,
  author = {Sabine Hensler and Claudia Kuehlbach and Bianka Kotkamp and Dario Frey and Margareta Maria Mueller},
  title = {A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery},
  journal = {American Journal of Bioscience and Bioengineering},
  volume = {9},
  number = {4},
  pages = {110-122},
  doi = {10.11648/j.bio.20210904.13},
  url = {https://doi.org/10.11648/j.bio.20210904.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20210904.13},
  abstract = {The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.},
 year = {2021}
}

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TY  - JOUR
T1  - A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery
AU  - Sabine Hensler
AU  - Claudia Kuehlbach
AU  - Bianka Kotkamp
AU  - Dario Frey
AU  - Margareta Maria Mueller
Y1  - 2021/08/11
PY  - 2021
N1  - https://doi.org/10.11648/j.bio.20210904.13
DO  - 10.11648/j.bio.20210904.13
T2  - American Journal of Bioscience and Bioengineering
JF  - American Journal of Bioscience and Bioengineering
JO  - American Journal of Bioscience and Bioengineering
SP  - 110
EP  - 122
PB  - Science Publishing Group
SN  - 2328-5893
UR  - https://doi.org/10.11648/j.bio.20210904.13
AB  - The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.
VL  - 9
IS  - 4
ER  -

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