Effect of microserum environment stimulation on extraction and biological function of colorectal cancer stem cells.
Cancer stem cells
Colon cancer tumor cells
Microserum environment
Tumor sphere enrichment
Journal
Discover. Oncology
ISSN: 2730-6011
Titre abrégé: Discov Oncol
Pays: United States
ID NLM: 101775142
Informations de publication
Date de publication:
28 Aug 2023
28 Aug 2023
Historique:
received:
16
05
2023
accepted:
22
08
2023
medline:
28
8
2023
pubmed:
28
8
2023
entrez:
28
8
2023
Statut:
epublish
Résumé
3D cancer stem cell (CSC) cultures are widely used as in vitro tumor models. In this study, we determined the effects of enriching HCT116 tumor spheres initially cultured in serum-free medium with different concentrations of serum, focusing on the effect of microserum environment stimulation on extraction and biological function of colorectal cancer stem cells (CCSCs). CCSCs were enriched in standard serum-free medium and serum-free medium with different concentrations of serum for 1 week. The expression of CSC-associated markers in CCSCs, and the presence and relative proportion of CSCs (CD133/CD44 cell sorting) were then determined to elucidate the effect of the microserum environment on the preservation of CSC-related features. Further, the tumorigenic capacity of CCSCs was evaluated in an immunodeficiency mouse model. Our data indicated that a significantly greater number of spheres with a greater size range and high viability without drastic alteration in biological and structural features, which maintained self-renewal potential after sequential passages were formed after serum supplementation. Real-time analysis showed that both serum spheres and serum-free spheres displayed similar expression patterns for key stemness genes. Serum spheres showed higher expression of the CSC surface markers CD133 and CD44 than did CSCs spheres cultured in serum-free medium. Adherent cultures in complete medium could adapt to the serum-containing microenvironment faster and showed higher proliferation ability. The addition of serum induced EMT and promoted the migration and invasion of serum globular cells. Compared with serum-free cells and adherent cells, serum spheres showed higher tumor initiation ability. Microserum environment stimulation could be an effective strategy for reliable enrichment of intact CCSCs, and a more efficient CSC enrichment method.
Sections du résumé
BACKGROUND
BACKGROUND
3D cancer stem cell (CSC) cultures are widely used as in vitro tumor models. In this study, we determined the effects of enriching HCT116 tumor spheres initially cultured in serum-free medium with different concentrations of serum, focusing on the effect of microserum environment stimulation on extraction and biological function of colorectal cancer stem cells (CCSCs).
METHODS
METHODS
CCSCs were enriched in standard serum-free medium and serum-free medium with different concentrations of serum for 1 week. The expression of CSC-associated markers in CCSCs, and the presence and relative proportion of CSCs (CD133/CD44 cell sorting) were then determined to elucidate the effect of the microserum environment on the preservation of CSC-related features. Further, the tumorigenic capacity of CCSCs was evaluated in an immunodeficiency mouse model.
RESULTS
RESULTS
Our data indicated that a significantly greater number of spheres with a greater size range and high viability without drastic alteration in biological and structural features, which maintained self-renewal potential after sequential passages were formed after serum supplementation. Real-time analysis showed that both serum spheres and serum-free spheres displayed similar expression patterns for key stemness genes. Serum spheres showed higher expression of the CSC surface markers CD133 and CD44 than did CSCs spheres cultured in serum-free medium. Adherent cultures in complete medium could adapt to the serum-containing microenvironment faster and showed higher proliferation ability. The addition of serum induced EMT and promoted the migration and invasion of serum globular cells. Compared with serum-free cells and adherent cells, serum spheres showed higher tumor initiation ability.
CONCLUSIONS
CONCLUSIONS
Microserum environment stimulation could be an effective strategy for reliable enrichment of intact CCSCs, and a more efficient CSC enrichment method.
Identifiants
pubmed: 37639070
doi: 10.1007/s12672-023-00779-z
pii: 10.1007/s12672-023-00779-z
pmc: PMC10462592
doi:
Types de publication
Journal Article
Langues
eng
Pagination
156Subventions
Organisme : Natural Science Foundation of Guizhou Province
ID : QianKeHe Support [2022]181
Organisme : Natural Science Foundation of Guizhou Province
ID : QianKeHe Basics -ZK[2023] Key 042
Organisme : Natural Science Foundation of Guiyang City
ID : [2022]4-3-10
Organisme : Natural Science Foundation of Guiyang City
ID : [2022]4-3-11
Organisme : Natural Science Foundation of Guiyang City
ID : [2022]4-3-2
Organisme : Project Foundation of Guizhou Administration of Traditional Chinese Medicine
ID : QZYYXG-2021-5
Organisme : National Natural Science Foundation of China
ID : 31660326
Organisme : National Natural Science Foundation of China
ID : 82160519
Informations de copyright
© 2023. Springer Science+Business Media, LLC.
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