PhD Scientific Days 2025

Pharmaceutical Sciences and Health Technologies III.

Development of a 3D HL-1 Cardiomyocyte Model System Using Self-Assembling Peptide

Előadó neve

Dr. Szász Zsófia

Neptun code

IKMTJT

Előadó munkahelye

Department of Genetics, Cell- and Immunobiology, Semmelweis University

Előadó telefonszáma

+36 1 210 2930/56236

Előadó e-mail címe

szasz.adrienn.zsofia@semmelweis.hu

Az előadás címe

Development of a 3D HL-1 Cardiomyocyte Model System Using Self-Assembling Peptide

Szerző(k) neve és munkahelye

Zsófia Szász¹, Petra Pőcze¹, Kata Nóra Enyedi², Dorina Lenzinger¹, Angéla Takács¹, László Kőhidai¹, Eszter Lajkó¹

1: Department of Genetics, Cell- and Immunobiology, Semmelweis University
2: Institute of Chemistry, Eötvös Loránd University

Bemutatás módja

Szóbeli

Szekció

Pharmaceutical Sciences and Health Technologies III.

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: Drug development begins in the preclinical phase with target identification and in vitro testing. There has been a growing focus on developing advanced 3D cell culture models that better replicate the complexity of in vivo tissues compared to traditional 2D monolayer cultures. These models not only allow for a more accurate understanding of drug mechanisms but also offer improved prediction of potential side effects (e.g. cardiotoxicity) before in vivo experiments.
Aim: The objective of this study was to develop a hydrogel-based 3D cardiomyocyte model using a self-assembling peptide (EAK-16-II). We also aimed to investigate (i) the effects of extracellular matrix (ECM) components - specifically fibronectin and laminin - (ii) and drug (daunorubicin – Dau) targeting peptide conjugates on spheroid size and cell viability.
Methods: HL-1 mouse cardiomyocytes were cultured in a self-assembling peptide hydrogel matrix for 3 or 6 days, with or without fibronectin or laminin. Spheroid size was measured using ImageJ from ZEISS Celldiscoverer7 images. Cardiomyocyte markers were analyzed via indirect immunocytochemistry using a Leica SP8 confocal microscope. To evaluate the model, 3-day spheroids were treated with Dau-peptide conjugates for 3 days, and viability was assessed with the CellTiter-Glo 3D assay.
Results: After 3 days, HL-1 cells formed spheroids within the hydrogel matrix with a mean diameter of 33±9.1 μm, increasing to 38±11.1 μm by day 6. The spheroids were cultured for up to a period of 6 days, and there was no decrease in cell viability. Adding fibronectin or laminin did not significantly alter spheroid size or viability. The HL-1 cells retained their cardiomyocyte markers, including α-actinin and caveolin, within the spheroids. Upon treatment with the Dau-based peptide conjugates, the 3D model exhibited greater sensitivity compared to 2D cultures, as evidenced by cell viability remaining above 50% in the 3D model, while viability in the 2D cultures stayed above 75%.
Conclusion: We successfully developed a 3D cardiomyocyte model using the self-assembling EAK-16-II peptide hydrogel. Although the resulting spheroids were relatively small, this platform shows promise for future development as a preclinical tool to evaluate cardiotoxic side effects of drug candidates prior to in vivo testing.
Funding: RRF-2.3.1-21–2022-00003, EKOP-2024-120

University

Semmelweis University

Supervisor

Eszter Lajkó

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

after finishing doctoral studies with absolutorium (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

6041

Start

16:15

End

16:30