a-quantitative-assessment-phagocytosis-allogeneic-xenogeneic

A Quantitative Assessment of the Phagocytosis of Allogeneic and Xenogeneic Erythrocytes by Rat Macrophages <em>In Vitro</em>

Xenogeneic cell transplantation often faces significant immune rejection, even in immunodeficient animal models. Among residual immune components, ...

Chinese Academy of Medical Sciences

Wuyi University

Fujian University of Traditional Chinese Medicine

Prometheus RegMed Tech Ltd

The increasing demand for disease modeling, preclinical drug testing, and long waiting lists for alternative organ substitutes has posed significant challenges to current limitations in organoid technology. Consequently, organoid technology has emerged as a cutting-edge tool capable of accurately recapitulating the complexity of actual organs in physiology and functionality. To bridge the gaps between basic research and pharmaceutical as well as clinical applications, efforts have been made to develop organoids from tissue-derived stem cells or pluripotent stem cells. These developments include optimizing starting cells, refining culture systems, and introducing genetic modifications. With the rapid development of organoid technology, organoid composition has evolved from single-cell to multi-cell types, enhancing their level of biomimicry. Tissue structure has become more refined, and core challenges like vascularization are being addressed actively. These improvements are expected to pave the way for the construction of organoid atlases, automated large-scale cultivation, and universally compatible organoid biobanks. However, major obstacles remain to be overcome before urgently proof-of-concept organoids can be readily converted to practical applications. These obstacles include achieving structural and functional summarily to native tissue, remodeling the microenvironment, and scaling up production. This review aims to summarize the status of organoid development and applications, highlight recent progress, acknowledge existing limitations and challenges, and provide insights into future advancements. It is expected that this will contribute to the establishment of a reliable, scalable, and practical platform for organoid production and translation, further promoting their use in the pharmaceutical industry and regenerative medicine.

Trends and challenges in organoid modeling and expansion with pluripotent stem cells and somatic tissue.

Hepatic stellate cells (HSCs) are liver-specific mesenchymal cells that play a crucial role in liver formation and regeneration, as well as in different pathological diseases. However, the limited source of primary HSCs (pHSCs) and the suboptimal functionality of induced HSCs (iHSCs) by existing methods restrict their application in biomedical modeling. We developed a de novo differentiation method to generate iHSCs under simulated liver microenvironment in vitro, thereby enhancing the function of the differentiated cells. These iHSCs exhibited key HSC functions, including the expression of α-smooth muscle actin, collagen, and the capability to store Vitamin A. RNA sequencing further revealed that the present iHSCs converged more closely to pHSCs with very similar transcriptional profile compared to the established conventional induction. Additionally, the novel HSC-specific marker genes, FBLN5, NID2, and SVEP1 were identified by RNA sequencing and gene expression assay. In conclusion, our novel differentiation approach enables the generation of iHSCs with phenotypic and functional traits similar to those of pHSCs. The generation of highly functional iHSCs may make it more feasible to accurately simulate the liver-specific multicellular microenvironments, thus providing new perspectives on the modeling of physiological regenerative processes and disease progression in the liver, as well as useful tools for creating of new therapeutic strategies.

Liver developmental microenvironment promotes iHSC generation from human iPSCs.

To address the limitations of conventional strategies for differentiation human induced pluripotent stem cells (hiPSCs) into homogeneous hepatocytes, a doxycycline-inducible GATA6-expressing hiPSC line was developed using the PiggyBac transposon system. This hiPSC line permitted temporal control of GATA6 expression, enabling synchronous differentiation into multilineage parenchymal and non-parenchymal liver cells within 18 days. The derived cells demonstrated mature hepatocyte-specific gene expression and sustained albumin secretion. This cell line provides a robust platform for generating functional 3D multilineage liver organoids, and offers new opportunities for mechanistic studies of liver disorders, disease modeling, and regenerative medicine.

Article	Year
Trends and challenges in organoid modeling and expansion with pluripotent stem cells and somatic tissue. PeerJ\| PubMed ID: 39619184	2024
Liver developmental microenvironment promotes iHSC generation from human iPSCs. Scientific reports\| PubMed ID: 40595388	2025
Generation of human induced pluripotent stem cells with inducible expression of GATA6（Liver cells from hiPSC-GATA6）. Stem cell research\| PubMed ID: 40753856	2025

Jian-Yun Ge

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