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AlgiMatrix™-Based 3D Cell Culture System as an In Vitro Tumor Model: An Important Tool in Cancer Research

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Cancer Chemoprevention

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1379))

Abstract

Routinely used two-dimensional cell culture-based models often fail while translating the observations into in vivo models. This setback is more common in cancer research, due to several reasons. The extracellular matrix and cell-to-cell interactions are not present in two-dimensional (2D) cell culture models. Diffusion of drug molecules into cancer cells is hindered by barriers of extracellular components in in vivo conditions, these barriers are absent in 2D cell culture models. To better mimic or simulate the in vivo conditions present in tumors, the current study used the alginate based three-dimensional cell culture (AlgiMatrix™) model, which resembles close to the in vivo tumor models. The current study explains the detailed protocols involved in AlgiMatrix™ based in vitro non-small-cell lung cancer (NSCLC) models.

The suitability of this model was studied by evaluating, cytotoxicity, apoptosis, and penetration of nanoparticles into the in vitro tumor spheroids. This study also demonstrated the effect of EphA2 receptor targeted docetaxel-loaded nanoparticles on MDA-MB-468 TNBC cell lines. The methods section is subdivided into three subsections such as (1) preparation of AlgiMatrix™-based 3D in vitro tumor models and cytotoxicity assays, (2) free drug and nanoparticle uptake into spheroid studies, and (3) western blot, IHC, and RT-PCR studies.

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Abbreviations

CSCs:

Cancer stem cells

DIO-NLC:

DIO dye-loaded NLC

EGF:

Epidermal growth factor

ECM:

Extracellular matrix

FGF:

Fibroblast growth factor

FBS:

Fetal bovine serum

IHC:

Immunohistochemistry

NSCLC:

Non-small-cell lung cancer

NLC:

Nanostructured lipid carriers

OCT:

Optimal cutting temperature

RT-PCR:

Reverse transcription-polymerase chain reaction

SEM:

Scanning electron microscopic

TNBC:

Triple-negative breast cancer

2D:

Two dimensional

3D:

Two dimensional

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Correspondence to Chandraiah Godugu or Mandip Singh .

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Godugu, C., Singh, M. (2016). AlgiMatrix™-Based 3D Cell Culture System as an In Vitro Tumor Model: An Important Tool in Cancer Research. In: Strano, S. (eds) Cancer Chemoprevention. Methods in Molecular Biology, vol 1379. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3191-0_11

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  • DOI: https://doi.org/10.1007/978-1-4939-3191-0_11

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3190-3

  • Online ISBN: 978-1-4939-3191-0

  • eBook Packages: Springer Protocols

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