PhD Scientific Days 2020

Pathology and Oncology I. Posters

Development and Characterization of 3 Dimensional In Vitro Model of Neuroendocrine Tumors

Előadó neve

Krokker, Lilla, MSc

Előadó munkahelye

Semmelweis Egyetem, Laboratóriumi Medicina Intézet

Előadó telefonszáma

702956789

Előadó e-mail címe

krkkr.lilla@gmail.com

Az előadás címe

Development and Characterization of 3 Dimensional In Vitro Model of Neuroendocrine Tumors

Szerző(k) neve és munkahelye

Krokker Lilla 1, 2, Szabó Borbála 1, 3 Németh Kinga1, Sarkadi Balázs3, Mészáros Katalin1, Patócs Attila1, 2, Butz Henriett1, 2

1Hereditary Endocrine Tumours Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary
2Department of Laboratory Medicine, Semmelweis University, Budapest, Hungary
32nd Department of Internal Medicine, Semmelweis University, Budapest, Hungary
4Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary

Szekció

Pathology and Oncology I. Posters

Language of the presentation

Hungarian

Section, first choice

Pathology and Oncology

Section, second choice

Health Sciences

Összefoglaló szövege

Introduction. In vitro two dimensional cell cultures are widely applied in cancer biology. However, there is a considerable difference among tumour cell behaviour of existing models and solid tumours because monolayer cell cultures do not represent the heterogeneity of human tumours reliably. Treatment options of endocrine tumours are limited and do not respond to radiation therapy either. Until now the information about 3D culturing of endocrine tumours is minimal.
Aim. Establishment of three-dimensional in vitro models of endocrine tumours and their comparison with monolayer cell cultures in order to identify novel pathomechanisms and potential new therapeutic targets.
Materials and methods. Induction of spheroid formation of pituitary tumours (GH3, RC-4B/C), adrenal carcinoma (H295R) and phaeochromocytoma (PC12) cell lines was performed by different methods: using serum-free defined medium, ultra-low attachment plate and matrigel. AlamarBlue assay was used to examine cell viability and proliferation. Live and dead cell ratios were determined by trypan blue exclusion assay. The effect mitotane treatment was assessed on H295R adrenocortical cells. Steroid hormone production was monitored by liquid chromatography coupled with tandem mass spectrometry. Inhibition of the SDHB enzyme in PC12 cells was done by itaconate and the glutaminase enzyme was inhibited by a selective inhibitor compound .
Results. While pituitary tumour cells were not able to form spheroids, H295R cells were capable of producing 3D structure using several conditions. Regarding PC12 cells spheroid induction was successful using SFDM. In 3D cultures H295R and PC12 cells further proliferated and kept their viability up to 10 days. H295R spheroid cell culture showed a higher tendency for cortisol production compared to monolayer cell culture. Significantly reduced hormone production capacity was identified after anti-proliferative effect of mitotane in spheroids. Regarding PC12 cells, the proliferation and viability upon itaconate and BPTES treatment of 3D and monolayer cultures showed difference.
Conclusion. Cell viability, growth characteristics and hormone production differed in 3D and 2D cultures and they showed different response to therapy as well. In vitro 3D models could be a useful model for neuroendocrine tumours mimicking solid tumour cell behaviour more reliably.

Additional Information

Supervisor: Dr. Butz Henriett
E-mail address: butz.henriett@med.semmelweis-univ.hu

Bemutatás módja

Szóbeli

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

4601

Start

11:35

End

11:38

Authors (legacy)

Krokker Lilla 1, 2, Szabó Borbála 1, 3 Németh Kinga1, Sarkadi Balázs3, Mészáros Katalin1, Patócs Attila1, 2, Butz Henriett1, 2

1Hereditary Endocrine Tumours Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary
2Department of Laboratory Medicine, Semmelweis University, Budapest, Hungary
32nd Department of Internal Medicine, Semmelweis University, Budapest, Hungary
4Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary