TY  - CONF
AU  - Stanković, Tijana
AU  - Dragoj, Miodrag
AU  - Stojkovska, J
AU  - Dinić, Jelena
AU  - Podolski-Renić, Ana
AU  - Obradović, B.
AU  - Pešić, Milica
PY  - 2020
UR  - https://www.eaa2020.org/abstracts
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4109
AB  - Introduction
Various three dimensional (3D) biomimicking human glioblastoma cell culture
models have been developed in the last decade. However, all these models have
limited duration of viable cell cultures, which is necessary for reliable drug response
studies. Therefore, we are developing a novel long-term 3D glioblastoma
biomimicking model system that would enable optimal drug testing at clinically
relevant duration.
Material and Methods
3D culture of human U87 glioblastoma cells in alginate microfibers was followed for
28 days under static and dynamic conditions. To characterize this culture, cell
growth and viability were assessed by trypan blue dye exclusion test every seven
days, until 28th day. At the same time points, cell morphology and aggregation
were analyzed by fluorescent and confocal microscopy upon calcein-AM/propidium
iodide staining. Drug testing was validated by comparing effects of two different
TMZ treatment modalities on cell viability, morphology, aggregation and resistancerelated
gene expression (MGMT and ABCB1). Specifically, effects of 3 treatments
with 100 μM TMZ, starting from day 7 (X=7), were compared between subsequent
treatment modality (day by day treatments, X+1) and protractive treatment
modality (every 7 day, X+7).
Results and Discussions
Within the newly established static 3D model system, U87 glioblastoma cells
remained viable up to 28 days. The number of cells increased over time, while the
cell death rate was low. At day 7, cells formed visible aggregates oriented to
microfiber periphery, towards the source of oxygen and nutrients. Importantly, both TMZ treatment modalities had the same effect on cell viability in the static longterm
3D culture. However, protractive treatment reduced the expression of MGMT
and ABCB1observed with subsequent treatments.
Further work will be focused on the development of dynamic long-term 3D cell
culture in perfusion bioreactor. In this advanced model system, U87 cells in alginate
microfibers will be exposed to continuous media flow for 28 days. Introduction of
media flow, as the tumor microenvironment factor, may promote different
glioblastoma phenotype making it more reliable in vitro model for drug testing.
Conclusion
Based on the results obtained so far, we can conclude that our model system could
be suitable for drug testing in glioblastoma gaining relevant results for future
clinical studies.
PB  - European Association for Cancer Research
C3  - EACR-AACR-ASPIC Basic and Translational Research Conference "Tumor Microenvironment"; 2020 Mar 02-04;Lisbon, Portugal
T1  - Validation of long-term 3D glioblastoma cell culture as a novel biomimicking model system for preclinical drug testing
SP  - P-303
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4109
ER  - 

@conference{
author = "Stanković, Tijana and Dragoj, Miodrag and Stojkovska, J and Dinić, Jelena and Podolski-Renić, Ana and Obradović, B. and Pešić, Milica",
year = "2020",
abstract = "Introduction
Various three dimensional (3D) biomimicking human glioblastoma cell culture
models have been developed in the last decade. However, all these models have
limited duration of viable cell cultures, which is necessary for reliable drug response
studies. Therefore, we are developing a novel long-term 3D glioblastoma
biomimicking model system that would enable optimal drug testing at clinically
relevant duration.
Material and Methods
3D culture of human U87 glioblastoma cells in alginate microfibers was followed for
28 days under static and dynamic conditions. To characterize this culture, cell
growth and viability were assessed by trypan blue dye exclusion test every seven
days, until 28th day. At the same time points, cell morphology and aggregation
were analyzed by fluorescent and confocal microscopy upon calcein-AM/propidium
iodide staining. Drug testing was validated by comparing effects of two different
TMZ treatment modalities on cell viability, morphology, aggregation and resistancerelated
gene expression (MGMT and ABCB1). Specifically, effects of 3 treatments
with 100 μM TMZ, starting from day 7 (X=7), were compared between subsequent
treatment modality (day by day treatments, X+1) and protractive treatment
modality (every 7 day, X+7).
Results and Discussions
Within the newly established static 3D model system, U87 glioblastoma cells
remained viable up to 28 days. The number of cells increased over time, while the
cell death rate was low. At day 7, cells formed visible aggregates oriented to
microfiber periphery, towards the source of oxygen and nutrients. Importantly, both TMZ treatment modalities had the same effect on cell viability in the static longterm
3D culture. However, protractive treatment reduced the expression of MGMT
and ABCB1observed with subsequent treatments.
Further work will be focused on the development of dynamic long-term 3D cell
culture in perfusion bioreactor. In this advanced model system, U87 cells in alginate
microfibers will be exposed to continuous media flow for 28 days. Introduction of
media flow, as the tumor microenvironment factor, may promote different
glioblastoma phenotype making it more reliable in vitro model for drug testing.
Conclusion
Based on the results obtained so far, we can conclude that our model system could
be suitable for drug testing in glioblastoma gaining relevant results for future
clinical studies.",
publisher = "European Association for Cancer Research",
journal = "EACR-AACR-ASPIC Basic and Translational Research Conference "Tumor Microenvironment"; 2020 Mar 02-04;Lisbon, Portugal",
title = "Validation of long-term 3D glioblastoma cell culture as a novel biomimicking model system for preclinical drug testing",
pages = "P-303",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4109"
}

Stanković, T., Dragoj, M., Stojkovska, J., Dinić, J., Podolski-Renić, A., Obradović, B.,& Pešić, M.. (2020). Validation of long-term 3D glioblastoma cell culture as a novel biomimicking model system for preclinical drug testing. in EACR-AACR-ASPIC Basic and Translational Research Conference "Tumor Microenvironment"; 2020 Mar 02-04;Lisbon, Portugal
European Association for Cancer Research., P-303.
https://hdl.handle.net/21.15107/rcub_ibiss_4109

Stanković T, Dragoj M, Stojkovska J, Dinić J, Podolski-Renić A, Obradović B, Pešić M. Validation of long-term 3D glioblastoma cell culture as a novel biomimicking model system for preclinical drug testing. in EACR-AACR-ASPIC Basic and Translational Research Conference "Tumor Microenvironment"; 2020 Mar 02-04;Lisbon, Portugal. 2020;:P-303.
https://hdl.handle.net/21.15107/rcub_ibiss_4109 .

Stanković, Tijana, Dragoj, Miodrag, Stojkovska, J, Dinić, Jelena, Podolski-Renić, Ana, Obradović, B., Pešić, Milica, "Validation of long-term 3D glioblastoma cell culture as a novel biomimicking model system for preclinical drug testing" in EACR-AACR-ASPIC Basic and Translational Research Conference "Tumor Microenvironment"; 2020 Mar 02-04;Lisbon, Portugal (2020):P-303,
https://hdl.handle.net/21.15107/rcub_ibiss_4109 .