Abstract
Objectives
To design novel 3D in vitro co-culture models based on the RGD-peptide-induced cell self-assembly technique.
Results
Multicellular spheroids from M-3 murine melanoma cells and L-929 murine fibroblasts were obtained directly from monolayer culture by addition of culture medium containing cyclic RGD-peptide. To reach reproducible architecture of co-culture spheroids, two novel 3D in vitro models with well pronounced core–shell structure from tumor spheroids and single mouse fibroblasts were developed based on this approach. The first was a combination of a RGD-peptide platform with the liquid overlay technique with further co-cultivation for 1–2 days. The second allowed co-culture spheroids to generate within polyelectrolyte microcapsules by cultivation for 2 weeks. M-3 cells (a core) and L-929 fibroblasts (a shell) were easily distinguished by confocal microscopy due to cell staining with DiO and DiI dyes, respectively.
Conclusions
The 3D co-culture spheroids are proposed as a tool in tumor biology to study cell–cell interactions as well as for testing novel anticancer drugs and drug delivery vehicles.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akasov R, Borodina T, Zaytseva E et al (2015) Ultrasonically assisted polysaccharide microcontainers for delivery of lipophilic antitumor drugs: preparation and in vitro evaluation. ACS Appl Mater Interface 7:16581–16589
Akasov R, Zaytseva-Zotova D, Burov S et al (2016) Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in vitro. Int J Pharm 506:148–157
Bartkowiak A, Brylak W (2006) Hydrogel microcapsules containing natural and chemically modified oligochitosans: mechanical properties and porosity. Polimery/Polymers 51:547–554
Brandner JM, Haass NK (2013) Melanoma’s connections to the tumour microenvironment. Pathology 45:443–452
Correia CR, Pirraco RP, Cerqueira MT et al (2016) Semipermeable capsules wrapping a multifunctional and self-regulated co-culture microenvironment for osteogenic differentiation. Sci Rep 6:21883
Costa EC, Gaspar VM, Coutinho P, Correia IJ (2014) Optimization of liquid overlay technique to formulate heterogenic 3D co-cultures models. Biotechnol Bioeng 111:1672–1685
Dittmer J, Leyh B (2015) The impact of tumor stroma on drug response in breast cancer. Semin Cancer Biol 31:3–15
Estrada MF, Rebelo SP, Davies EJ et al (2016) Modelling the tumour microenvironment in long-term microencapsulated 3D co-cultures recapitulates phenotypic features of disease progression. Biomaterials 78:50–61
Fang X, Sittadjody S, Gyabaah K et al (2013) Novel 3D co-culture model for epithelial-stromal cells interaction in prostate cancer. PLoS One 8:e75187
Flach EH, Rebecca VW, Herlyn M et al (2011) Fibroblasts contribute to melanoma tumor growth and drug resistance. Mol Pharm 8:2039–2049
Hickman J, Graeser R, de Hoogt R et al (2014) Three-dimensional models of cancer for pharmacology and cancer cell biology: capturing tumor complexity in vitro/ex vivo. Biotechnol J 9:1115–1128
Horie M, Saito A, Yamaguchi Y et al (2015) Three-dimensional co-culture model for tumor–stromal interaction. J Vis Exp 96:e52469
Hsiao AY, Tung Y-C, Qu X et al (2012) 384 hanging drop arrays give excellent Z-factors and allow versatile formation of co-culture spheroids. Biotechnol Bioeng 109:1293–1304
Ivanov DP, Parker TL, Walker DA et al (2015) In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment. J Biotechnol 205:3–13
Jones TS, Holland EC (2012) Standard of care therapy for malignant glioma and its effect on tumor and stromal cells. Oncogene 31:1995–2006
Mehta G, Hsiao AY, Ingram M et al (2012) Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release 164:192–204
Metzger W, Sossong D, Bächle A et al (2011) The liquid overlay technique is the key to formation of co-culture spheroids consisting of primary osteoblasts, fibroblasts and endothelial cells. Cytotherapy 13:1000–1012
No DY, Lee S-A, Choi YY et al (2012) Functional 3D human primary hepatocyte spheroids made by co-culturing hepatocytes from partial hepatectomy specimens and human adipose-derived stem cells. PLoS One 7:e50723
Sutherland RM, McCredie JA, Inch WR (1971) Growth of multicell spheroids in tissue culture as a model of nodular carcinomas. J Natl Cancer Inst 46:113–120
Zaytseva-Zotova D, Balysheva V, Tsoy A et al (2011a) Biocompatible smart microcapsules based on chitosan-poly(vinyl alcohol) copolymers for cultivation of animal cells. Adv Eng Mater 13:493–500
Zaytseva-Zotova DS, Udartseva OO, Andreeva ER et al (2011b) Polyelectrolyte microcapsules with entrapped multicellular tumor spheroids as a novel tool to study the effects of photodynamic therapy. J Biomed Mater Res 97B:255–262
Supporting Information
Supplementary Fig. 1—Spheroid size distributions and micrographs of spheroids generated from M-3 and L-929 cells at various cell count ratios.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
10529_2016_2218_MOESM1_ESM.tif
Supplementary Fig. 1. Spheroid size distributions and micrographs of spheroids generated from M-3 and L-929 cells at various cell count ratios. To form spheroids, the cells were incubated with 50 µM of cyclo-RGDfK(TPP) for 72 h. Scale bar 100 µm. (TIFF 6955 kb)
Rights and permissions
About this article
Cite this article
Akasov, R., Gileva, A., Zaytseva-Zotova, D. et al. 3D in vitro co-culture models based on normal cells and tumor spheroids formed by cyclic RGD-peptide induced cell self-assembly. Biotechnol Lett 39, 45–53 (2017). https://doi.org/10.1007/s10529-016-2218-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-016-2218-9