[3S06e] 16. Cell and Tissue Engineering for Biomedical Process Innovation - Part 3

5:50 PM - 6:10 PM JST(8:50 AM - 9:10 AM UTC)

[3CTBP-13] Explanted human liver derived extracellular matrix for the establishment of patient derived liver organoids

○Tanveer Ahamd Mir^1,2, Fayrouz Mustafa^1,2, Eman A. Alshehri¹, Dimitri A. Raptis^1,2, Dieter C. Broering^1,2, Abdullah M. Assiri^1,2 (1. King Faisal Specialist Hospital & Research Centre, Riyadh, KSA (Saudi Arabia), 2. College of Medicine, Alfaisal University, Riyadh 11211, KSA (Saudi Arabia))

Keywords:

human liver,extracellular matrix,organoids

Background: Functional liver organoid models have been instrumental in simulating molecular mechanisms responsible for many cellular processes and disease states¹. However, established biomaterial substrates used for preparing and controlling 3D microenvironment largely rely on expensive and poorly defined tumor-cell-derived extracellular matrices (ECM)². Matrigel based ECM display significant batch variability, reproducibility issues and thus limit its possible applications in translational regenerative medicine. Thus, there has been considerable interest in identifying mechanically tunable and biocompatible alternative ECM-mimetics for the growth and expansion of liver organoids^3-4. Here we investigated the ability of explanted human liver-derived extracellular matrix particles to model the 3D microenvironment in vitro. The proposed material supported viability and growth of the human organoids with defined morphologies. Experimental Methods: Explanted human liver biopsies were collected post-surgery at KFSHRC, Riyadh (Protocol No: 2240025). Tissues were decellularized via freeze–thaw cycles and Triton X-100 treatment. Pregel dECM solution was prepared by dissolving the lyophilized dECM pieces with pepsin. The final hydrogel was prepared by mixing the dECM particles with different weight ratios. Proteins were analyzed using label-free liquid chromatography–tandem mass spectrometry, followed by pathway and network analysis. Finally, patient biopsy derived organoids were established ⁵. Results and Discussion: Hematoxylin and eosin , and IHC staining of liver tissue before and after decellularization showed the absence of cellular components and maintenance of liver-specific ECM components. The proposed gel was used for the subsequent culture and encapsulation of patient biopsy derived organoids. The organoids were maintained for time points, and images were captured. Conclusions: Our preliminary results indicated that the concentration of proposed biomaterials play an important role in growth and viability of the organoids. Furthermore, the hydrophilic and mechanical properties of the dECM were enhanced when enriched with natural polymers. The data suggest that the proposed approach is a promising alternative biomaterial that can be used for experimental organoid studies and high-throughput drug screening.
References
Mir et al., Int J Bioprint . 2023 (3),714.
Arai, K. F.Obonai , .. T.A. Mir et al.,Engineered Regeneration 7, 1-11,2026.
Obeid D.A, Mir, T.A et al. Biomedicines 2024, 12, 446
Wani S. I. Mir, T.A. Mir, Nakamura M, et al. Bioprinting 2024, 42 , e00355.
Hu et al, Cell . 2018, 29;175(6),1591-1606.e19.