The Impact of Astrocytes and Endothelial Cells on Glioblastoma Stemness Marker Expression in Multicellular Spheroids

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dc.contributor.authorNakod, Pinaki S.
dc.contributor.authorKim, Yonghyun
dc.contributor.authorRao, Shreyas S.
dc.contributor.otherUniversity of Alabama Tuscaloosa
dc.date.accessioned2023-09-28T19:11:26Z
dc.date.available2023-09-28T19:11:26Z
dc.date.issued2021
dc.description.abstractIntroduction Glioblastoma multiforme (GBM), the most common primary brain tumor in adults, is extremely malignant and lethal. GBM tumors are highly heterogenous, being comprised of cellular and matrix components, which contribute to tumor cell invasion, cancer stem cell maintenance, and drug resistance. Here, we developed a heterotypic 3D spheroid model integrating GBM cells with astrocytes and endothelial cells (ECs) to better simulate the cellular components of the tumor microenvironment and investigate their impact on the stemness marker expression of GBM cells, which has not been previously investigated. Methods We used U87 GBM cells, C8-D1A mouse astrocytes, and human umbilical vein ECs to construct co- and tri-culture spheroid models in low-attachment U-well plates. We characterized the expression of known stemness markers NESTIN, SOX2, CD133, NANOG, and OCT4 in these models and compared it to respective mixed monoculture spheroids (control) using qRT-PCR and immunostaining. Results We incorporated GBM cells and astrocytes/ECs in 1:1, 1:2, 1:4, and 1:9 ratio and observed spontaneous self-assembled spheroids in all coculture conditions. We observed changing spheroid size dynamics over 7 days and an increased expression in stemness markers in GBM-astrocyte and GBM-EC coculture spheroids in 1:4 and 1:9 coculture conditions, respectively. In a triculture model employing GBM cells, astrocytes, and ECs in a 1:4:9 ratio, we found an increased expression of all the stemness markers. Conclusions We elucidated the impact of astrocytes and ECs on GBM stemness marker expression. This multicellular spheroid model may provide an important tool for investigating the crosstalk between cell types in GBM.en_US
dc.format.mediumelectronic
dc.format.mimetypeapplication/pdf
dc.identifier.citationNakod, P. S., Kim, Y., & Rao, S. S. (2021). The Impact of Astrocytes and Endothelial Cells on Glioblastoma Stemness Marker Expression in Multicellular Spheroids. In Cellular and Molecular Bioengineering (Vol. 14, Issue 6, pp. 639–651). Springer Science and Business Media LLC. https://doi.org/10.1007/s12195-021-00691-y
dc.identifier.doi10.1007/s12195-021-00691-y
dc.identifier.orcidhttps://orcid.org/0000-0001-6344-1258
dc.identifier.orcidhttps://orcid.org/0000-0001-7649-0171
dc.identifier.urihttps://ir.ua.edu/handle/123456789/10978
dc.languageEnglish
dc.language.isoen_US
dc.publisherSpringer
dc.subjectBrain cancer
dc.subjectCancer stem cells
dc.subjectCoculture
dc.subjectTriculture
dc.subjectSpheroids
dc.subjectGLIOMA-CELLS
dc.subjectMATRIX
dc.subjectMODEL
dc.subjectCHEMOTHERAPY
dc.subjectRESISTANCE
dc.subjectINVASION
dc.subjectDORMANCY
dc.subjectSIZE
dc.subjectCell & Tissue Engineering
dc.subjectBiophysics
dc.subjectCell Biology
dc.subjectEngineering, Biomedical
dc.titleThe Impact of Astrocytes and Endothelial Cells on Glioblastoma Stemness Marker Expression in Multicellular Spheroidsen_US
dc.typeArticle
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