TY - JOUR
T1 - Assessment of intratumoral heterogeneity in isolated human primary high-grade glioma
T2 - Cluster of differentiation 133 and cluster of differentiation 15 double staining of glioblastoma subpopulations
AU - Faried, Ahmad
AU - Widowati, Wahyu
AU - Rizal, Rizal
AU - Bolly, Hendrikus M.B.
AU - Halim, Danny
AU - Widodo, Wahyu S.
AU - Wibowo, Satrio H.B.
AU - Noverina, Rachmawati
AU - Tjahjono, Firman P.
AU - Arifin, Muhammad Z.
N1 - Funding Information:
A. Faried supported by the Grants-in-Aid from Universitas Padjadjaran, West Java, Indonesia.
Publisher Copyright:
© 2021 Ahmad Faried, Wahyu Widowati, Rizal Rizal, Hendrikus M.B. Bolly, Danny Halim, Wahyu S. Widodo, Satrio H.B. Wibowo, Rachmawati Noverina, Firman P. Tjahjono, Muhammad Z. Arifin.
PY - 2021
Y1 - 2021
N2 - BACKGROUND: Gliomas are the most common primary brain tumors, representing 50–60% of malignant primary brain tumors. Gliomas are highly heterogeneous with marked inter-and intratumoral diversity. Gliomas heterogeneity is a challenging issue in the development of personalized treatment. The simplest method for studying heterogeneity is using ex vivo cell cultures; in our case, the cell lines were isolated from patient with glioblastomas. AIM: Here, we reported distinct cell subpopulations heterogeneity in glioblastoma cells. METHODS: Human glioblastoma cells isolation is conducted by enzymatic method with combination of collagenase I, hyaluronidase, and trypsin enzyme in proportional amount from patient. Immunostaining was performed to assess glial fibrillary acidic protein (GFAP), Ki-67, isocitrate dehydrogenase-1 (IDH-1) status, and program death ligand-1 (PD-L1) expression. Primary glioblastoma cell line was characterized by flow cytometry (fluorescence-activated cell sorting) analysis based on cluster of differentiation (CD) 133 and CD15 marker expression. U87MG and CGNH-89 cell lines were used as control. Distinct subpopulation analysis was performed by double staining of CD133 and CD15 in isolated primary glioblastoma cell line and its comparative control cells. RESULTS: Our isolated glioblastoma cells morphology was adherent cells which were able to form spheres depending on environment. Immunostaining confirmed GFAP, Ki-67, IDH-1 mutants, and PD-L1 expression. Our isolated glioblastoma cells expressed CD133 and CD15, coexpressed CD133/CD15 in different patterns. The highest subpopulation in primary glioblastoma was CD133+/CD15+. CONCLUSION: Glioblastoma cells can be isolated using enzymatic methods. Isolated glioblastoma cells consist of four different subpopulations distinguished by CD133/CD15 double staining. Intratumoral heterogeneity exists and directly or indirectly depends on their microenvironment.
AB - BACKGROUND: Gliomas are the most common primary brain tumors, representing 50–60% of malignant primary brain tumors. Gliomas are highly heterogeneous with marked inter-and intratumoral diversity. Gliomas heterogeneity is a challenging issue in the development of personalized treatment. The simplest method for studying heterogeneity is using ex vivo cell cultures; in our case, the cell lines were isolated from patient with glioblastomas. AIM: Here, we reported distinct cell subpopulations heterogeneity in glioblastoma cells. METHODS: Human glioblastoma cells isolation is conducted by enzymatic method with combination of collagenase I, hyaluronidase, and trypsin enzyme in proportional amount from patient. Immunostaining was performed to assess glial fibrillary acidic protein (GFAP), Ki-67, isocitrate dehydrogenase-1 (IDH-1) status, and program death ligand-1 (PD-L1) expression. Primary glioblastoma cell line was characterized by flow cytometry (fluorescence-activated cell sorting) analysis based on cluster of differentiation (CD) 133 and CD15 marker expression. U87MG and CGNH-89 cell lines were used as control. Distinct subpopulation analysis was performed by double staining of CD133 and CD15 in isolated primary glioblastoma cell line and its comparative control cells. RESULTS: Our isolated glioblastoma cells morphology was adherent cells which were able to form spheres depending on environment. Immunostaining confirmed GFAP, Ki-67, IDH-1 mutants, and PD-L1 expression. Our isolated glioblastoma cells expressed CD133 and CD15, coexpressed CD133/CD15 in different patterns. The highest subpopulation in primary glioblastoma was CD133+/CD15+. CONCLUSION: Glioblastoma cells can be isolated using enzymatic methods. Isolated glioblastoma cells consist of four different subpopulations distinguished by CD133/CD15 double staining. Intratumoral heterogeneity exists and directly or indirectly depends on their microenvironment.
KW - Cluster of differentiation 133
KW - Cluster of differentiation 15
KW - Glioblastoma
KW - Glioma stem cells
KW - Subpopulations
UR - http://www.scopus.com/inward/record.url?scp=85101183161&partnerID=8YFLogxK
U2 - 10.3889/oamjms.2021.5516
DO - 10.3889/oamjms.2021.5516
M3 - Article
AN - SCOPUS:85101183161
SN - 1857-5749
VL - 9
SP - 87
EP - 94
JO - Open Access Macedonian Journal of Medical Sciences
JF - Open Access Macedonian Journal of Medical Sciences
ER -