Synergistic role of three dimensional niche and hypoxia on conservation of cancer stem cell phenotype
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Date
2016
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Abstract
Hypoxia is a pathalogical condition in which tissues are deprived of adequate oxygen supply. The hypoxia effect on tumors has a critically important role on maintenance of cancer stem cell phenotype. The aim of this study is to investigate the effects of hypoxia on cancer stem cells on three dimensional (3D) in vitro culture models. Osteosarcoma stem cells characterized by CD133 surface protein were isolated from osteosarcoma cell line (SaOS-2) by magnetic-activated cell sorting (MACS) technique. Isolated CD133+ and CD133− cells were cultivated under hypoxic (1% O2) and normoxic conditions (21% O2) for 3 days. For the 3D model, bacterial cellulose scaffold was used as the culture substrate. 3D morphologies of cells were examined by scanning electron microscopy (SEM); RT-PCR and immunocytochemistry staining were used to demonstrate conservation of the cancer stem cell phenotype in 3D environment under hypoxic conditions. Cell viability was shown by MTT assay on 3. and 7. culture days. This study is seen as an introduction to develop a 3D hypoxic cancer stem cell based tumor model to study CSC behavior and tumor genesis in vitro. © 2016
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Keywords
AC133 Antigen , Cell Count , Cell Hypoxia , Cell Line, Tumor , Cell Shape , Humans , Immunohistochemistry , Immunomagnetic Separation , Neoplastic Stem Cells , Osteosarcoma , Phenotype , Real-Time Polymerase Chain Reaction , RNA, Messenger , Stem Cell Niche , CD133 antigen , cellulose , membrane protein , molecular scaffold , CD133 antigen , messenger RNA , Article , cancer stem cell , cell hypoxia , cell selection , cell separation , cell structure , cell viability , conservation biology , controlled study , human , human cell , immunocytochemistry , in vitro study , magnetic activated cell sorting , MTT assay , phenotype , reverse transcription polymerase chain reaction , Saos 2 cell line , scanning electron microscopy , stem cell niche , tumor microenvironment , cancer stem cell , cell count , cell hypoxia , cell shape , genetics , immunohistochemistry , immunomagnetic separation , metabolism , osteosarcoma , pathology , phenotype , real time polymerase chain reaction , tumor cell line