PS4.5: Application of aptamer-cellulose composite hydrogels in the cultivation of engineered cholangiocyte organoids.

作者：Guobin Huang, Guoliang Tang, Xuechun Zhao, Zipei Wang, Guang Yang, Bo Yang, Zhishui Chen · 发表于：Transplantation · 年份：2025 · DOI：10.1097/01.tp.0001170356.60660.64 · 研究领域：Liver physiology and pathology、Pediatric Hepatobiliary Diseases and Treatments、Pancreatic function and diabetes

Introduction: Nowadays, a major limitation for organoids culturing is the dependence on Matrigel. Matrigel’s undefined makeup and batch variations as well as its animal source limit its use in human therapies. Consequently, researchers were exploring hydrogels with clearer compositions as replacements. Recent work has used programmable DNA-based hydrogels to grow organoids. However, integrating functional DNA aptamers into hydrogels for organoid culture hasn’t been reported. This study aimed to develop a hydrogel made from cellulose and equipped with aptamers to culture cholangiocyte organoids (COs). Methods: EpCAM-CELAPT MINI dual-aptamer chains were synthesized by rolling circle amplification (RCA), which were conjugating with Nanocellulose fibrils (CNF) via CaCl2 to fabricate Aptamer-functionalized CNF (Apt/CNF) hydrogels. Material properties were characterized by scanning electron microscope (SEM), X-Ray Diffraction (XRD), Fourier transform infrared (FT-IR), and rheological testing. COs were derived from human bile duct tissues. Organoids grown in Apt-CNF hydrogel versus Matrigel controls were evaluated for: (1) viability (Calcein-AM/PI), (2) marker expression (PCR/immunofluorescence for EpCAM/CK19/CK18), and (3) function (ELISA for albumin/bile acid secretion). Results: Firstly, the SEM micrograph revealed a porous structure of hydrogel that facilitated cell adhesion and growth. The XRD analysis and the FTIR spectrum confirmed the presence of phosphorus, which indicated ...