Abstract
Novel experimental conditions of cancer cell line culture have evolved throughout the recent years, with significantly growing interest in xeno-free, serum-free and three-dimensional culture variants. The choice of proper culture media may enable to mimic tumor microenvironment and promotion of cancer stem cells proliferation. To assess whether stem-like phenotype inducing media may be applied in renal cancer stem cell research, we performed a widespread screening of 13 cell culture media dedicated for mesenchymal cells, stem cells as well as mesenchymal stem cells. We have also screened extracellular matrix compounds and selected optimal RCC 3D—ECM supported culture model. Our results revealed that 786-O as well as HKCSCs cell line cultures in xeno-free media (NutriStem/StemXvivo) and laminin coated plates provide a useful tool in RCC cancer biology research and at the same time enable effective drug toxicity screening. We propose bio-mimic 3D RCC cell culture model with specific low-serum and xeno-free media that promote RCC cell viability and stem-like phenotype according to the tested genes encoding stemness factors including E-cadherin, N-cadherin, HIF1, HIF2, VEGF, SOX2, PAX2 and NESTIN.
Highlights
The vast majority of molecular cancer studies are conducted using immortalized cell lines cultured as two-dimensional (2D) monolayer on polystyrene surface, which does not reflect tumor in vivo structure and constrain stem-like phenotype (Bielecka et al 2016; Cattin et al 2018; Khawar et al 2018; Boghaert et al 2017)
We propose biomimic 3D RCC cell culture model with specific lowserum and xeno-free media that promote RCC cell viability and stem-like phenotype according to the tested genes encoding stemness factors including E-cadherin, N-cadherin, HIF1, HIF2, VEGF, SOX2, PAX2 and NESTIN
In particular it was shown that cancer stem cells that are found in 3D cultures express stemness genes encoding SRY-Box 2: SOX2, Octamer-binding transcription factor 4: OCT-4, or Homeobox transcription factor Nanog, and proangiogenic growth factors like vascular endothelial growth factor: VEGF (Bielecka et al 2016)
Summary
The vast majority of molecular cancer studies are conducted using immortalized cell lines cultured as two-dimensional (2D) monolayer on polystyrene surface, which does not reflect tumor in vivo structure and constrain stem-like phenotype (Bielecka et al 2016; Cattin et al 2018; Khawar et al 2018; Boghaert et al 2017). In particular it was shown that cancer stem cells that are found in 3D cultures express stemness genes encoding SRY-Box 2: SOX2, Octamer-binding transcription factor 4: OCT-4, or Homeobox transcription factor Nanog, and proangiogenic growth factors like vascular endothelial growth factor: VEGF (Bielecka et al 2016).
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