PhD Scientific Days 2024

Poster Session Q - Pathological and Oncological Sciences 2.

Transition from 2D to 3D Cell Culturing in Breast Cancer Research

Előadó neve

Ms. Moldvai, Dorottya

Neptun code

JQCZNN

Előadó munkahelye

Department of Pathology and Experimental Cancer Research

Előadó telefonszáma

06203291961

Előadó e-mail címe

moldvai.dorottya@gmail.com

Az előadás címe

Transition from 2D to 3D Cell Culturing in Breast Cancer Research

Szerző(k) neve és munkahelye

Dorottya Moldvai¹, Dániel Sztankovics¹, Gábor Petővári¹, Titanilla Dankó¹, Ildikó Krencz¹, Risa Miyaura¹, Rebeka Gelencsér¹, Anna Sebestyén¹

1: Department of Pathology and Experimental Cancer Research

Bemutatás módja

Poszter

Szekció

Poster Session Q - Pathological and Oncological Sciences 2.

Language of the presentation

Hungarian

Preferred session

Pathological and Oncological Sciences

Összefoglaló szövege

Introduction

The objectives of both the FDA and EMA to phase out animal testing have generated a pressing requirement for alternative testing approaches. In vitro models are emerging as substitutes for animal experimentation. 3D models are increasingly favored due to their enhanced precision and reproducibility, addressing the growing necessity for more efficient testing methodologies.

Aims

3D bioprinted tissue-mimetic structures’ (TMSs) unique attributes call for the validation of analytical techniques applied to traditional cell cultures. Our study endeavors to verify the effectiveness of cell proliferation assays on 3D bioprinted structures, investigate the tissue formation process and comparing the metabolic features and mTOR inhibitor sensitivity to traditional 2D cell cultures.

Methods

Using a 3D bioprinter (GeSiM Bioscaffolder 3.2.), the printing process (including bioink selection, structural design, and printing parameters), as well as maintenance protocol were established and refined for breast cancer cell line T47D. Cell proliferation assays (Alamar Blue and Sulphorhodamine B) and sample preparation methods suitable for analytical techniques (such as IHC, Wes Simple, and proteomics) have been developed and validated for the printed TMSs. mTOR inhibitor sensitivity and metabolic characterization studies of TMSs were also performed.

Results

TMSs were maintained for 21 days, and during this time, tissue formation was detectable after the 7th day. This observation was confirmed through diverse proliferation assays and histological analysis of paraffin-embedded tissue sections. Decreased mTOR-pathway related protein expression (pS6; pAkt) and mTOR activity (pS6/S6 and pAkt/Akt ratio) was observed in TMSs compared to 2D cell cultures. mTOR inhibitor sensitivity (rapamycin, 50 ng/ml; ipatasertib, 1 uM) also decreased in TMSs which was related to the former observed mTOR activity changes.

Conclusion

Our findings highlight the influence of tissue culture conditions on metabolic characteristics, thereby impacting in vitro study outcomes. It is imperative to employ in vitro test systems during drug evaluations that closely mimic in situ reactions, thus enhancing the success rate of drug developments, leading to potential savings in both time and money.

Supported by: TKP2021-EGA-24, NKFI-K-142799, EFOP-3.6.3-VEKOP-16-2017-00009.

University

Semmelweis University

Supervisor

Dr. Sebestyén Anna, DSc

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

6117

Start

16:35

End

16:38