Enrichment of cancer stem cells by agarose multi-well dishes and 3D spheroid culture

Cell and Tissue Research
Xiaoling GuoRenshan Ge

Abstract

As the theory of cancer stem cells (CSCs) is maturing, CSC-targeted therapy is emerging as an important therapeutic strategy and seeking the ideal method for rapid enrichment and purification of CSCs has become crucial. So far, based on the known CSC phenotypes and biological characteristics, the methods for enrichment CSCs mainly include low adhesion culture, low oxygen culture, chemotherapy drug stimulation and side population (SP) sorting but these methods cannot realize quick enrichment of the desired CSCs. Herein, we adopt a novel method that efficiently enriches a certain amount of CSCs through agarose multi-well dishes using rubber micro-molds to make cancer cells into cell spheroids (3D). These 3D cancer cell spheroids in the proportions of expression of CSC biomarkers (single stain of CD44, CD44v6 and CD133 or double stain of both CD44 and CD133) were significantly higher than those of the conventional adherent culture (2D) using flow cytometry analysis. In addition, the expression levels of stemness transcription factors such as OCT4, NANOG and SOX2 in 3D were also significantly higher than that in 2D through Western blot (WB) and quantitative polymerase chain reaction (qPCR) assays. In addition, the CSCs in 3D could ...Continue Reading

References

Mar 12, 2003·Proceedings of the National Academy of Sciences of the United States of America·Muhammad Al-HajjMichael F Clarke
Feb 6, 2004·Cancer Immunology, Immunotherapy : CII·Karl-Heinz HeiderGerd Munzert
Jun 23, 2004·Critical Reviews in Oncology/hematology·Stewart Sell
Dec 3, 2005·Cancer Research·Anne T CollinsNorman J Maitland
Nov 24, 2006·Nature·Lucia Ricci-VitianiRuggero De Maria
Jan 19, 2008·Nature·Tobias SchattonMarkus H Frank
Nov 5, 2008·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Lei DuQuan Chen
Feb 11, 2009·Cell·Jeong Beom KimHans R Schöler
May 6, 2009·Stem Cells·Shigeo TakaishiTimothy C Wang
Jul 8, 2009·Methods in Molecular Biology·Chenwei LiDiane M Simeone
Jul 21, 2009·International Journal of Cancer. Journal International Du Cancer·Weizhu ZhuEdward R Sauter
Mar 23, 2010·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·Véronique Orian-Rousseau
Feb 8, 2011·CA: a Cancer Journal for Clinicians·Ahmedin JemalDavid Forman
Apr 12, 2011·International Journal of Cancer. Journal International Du Cancer·Ilona KryczekWeiping Zou
Nov 10, 2011·Lab on a Chip·Alireza SalmanzadehRafael V Davalos
Jan 18, 2012·The International Journal of Biochemistry & Cell Biology·Marco A Velasco-VelázquezRichard G Pestell
Feb 7, 2012·Proceedings of the National Academy of Sciences of the United States of America·Katia MeirellesXiaolong Wei
Jun 12, 2012·Oral Oncology·Chih-Yen ChienChang-Han Chen
Jun 26, 2012·Cell and Tissue Research·Henry PageEmmanuel G Reynaud
Jun 27, 2012·Cell Cycle·Xingwang HuChiang J Li
Mar 13, 2014·Journal of Biotechnology·Xiao-xi XuXiao-jun Ma
Jan 13, 2015·Differentiation; Research in Biological Diversity·Sudhakara Rao PattabhiThomas C S Keller
Mar 5, 2017·Seminars in Cancer Biology·Claudia PeitzschAnna Dubrovska
Oct 27, 2017·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Xiong JinHyunggee Kim
Nov 2, 2017·Oncotarget·Cheng-Liang ZhangDong Liu
Nov 8, 2017·Theranostics·Gang ZhouLiang Qiao

❮ Previous
Next ❯

Methods Mentioned

BETA
flow cytometry
FCS
immunoprecipitation assay
Assay
PCR

Software Mentioned

Image J
GraphPad
GraphPad Prism
FCS Express

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.