Patient-derived organoids (PDOs) are revolutionizing cancer research, serving as invaluable models for tumor biology and therapeutic screening. The fidelity and applicability of these organoids are fundamentally shaped by the tissue dissociation techniques employed, namely mechanical dissociation and enzymatic digestion. This comprehensive review delves into the nuances of these two methods, scrutinizing their effects on solid tumor organoid properties, including stemness, heterogeneity, long-term culturing. We discuss the advantages and limitations of each technique, with a focus on their impact on tumor microenvironment preservation, their application in drug screening and cancer modeling. Moreover, we examine how recent technological breakthroughs have bolstered the efficiency and scalability of organoid production through these methods. Our analysis is designed to assist researchers in choosing the optimal tissue dissociation strategy for their research objectives and to fuel the evolution of organoid-based cancer models.

Objective: In this study, we investigated the cause of the AngII dose elevation in aqueous humor of primary open-angle glaucoma (POAG) patients. Methods: Enzyme-linked immunosorbent assay (ELISA), western blotting were used to detect concentration of Angiotensin Converting Enzyme 2 (ACE2) and Prolylcarboxypeptidase (PRCP). AngII and AngII + Recombinant PRCP were injected into anterior chamber of mouse eye. Mouse Intraocular pressure (IOP) was measured every week, mouse eye sections were conducted Hematoxylin-and-Eosin (H&E) staining, Masson’ staining and Immunofluorescence staining. Western blotting and Immunofluorescence staining assays to detected fibrosis of trabecular meshwork cells. Mass spectrometry was used to identify proteins of aqueous humor. Results: PRCP dose are decreased in aqueous humor of POAG patients. There is a negative correlation between PRCP and AngII levels in aqueous humor and between PRCP levels and the IOP. PRCP treatment reverses fibrosis of trabecular meshwork (TM) and prevents IOP elevation induced by AngII. Exogenous PRCP rescues fibrosis induced by AngII in HTMCs. Proteome profiling detected 502 differentially expressed proteins. Conclusion: Our study found PRCP dose was decreased in POAG patients’ aqueous humor, and it might cause high level of AngII. Restoration of PRCP rescued fibrosis of TM cells and ameliorated IOP in AngII treatment mouse.

Objective: In this study, we investigated the protective effect of Metformin on fibrosis of trabecular meshwork cells induced by TGFβ2. Methods: Transformed and primary human trabecular meshwork cells (HTMCs) were treated with TGFβ2 or Metformin alone or combination, western blotting and immunofluorescence staining assays to detect autophagy activity and fibrotic proteins expression levels. TGFβ2 or Metformin alone or combination were injected into the anterior chamber of mouse eye. Mouse intraocular pressure (IOP) was measured every week, mouse eye sections were conducted immunofluorescence staining to analyze Col1 and Col3 expression. pSmad3 level and localization to evaluate TGFβ/Smad3 pathway activity. Chloroquine phosphate was used to block autophagy-lysosome pathway. Results: Metformin activates autophagy of HTMCs in a dose dependent manner and efficiently ameliorates TMCs fibrosis induced by TGFβ2 in vitro and in mouse model, and decreased elevated IOP caused by TGFβ2. Metformin promotes fibrotic proteins degradation through the autophagy-lysosome pathway. Conclusion: Our study found Metformin could alleviates fibrosis of HTMCs induced by TGFβ2 and decreased elevated IOP in mouse model.

Approximately 60% of glaucoma patients have a family history. Family‐based studies indicate that relatives of glaucoma patients have a 10‐fold higher risk of developing glaucoma compared to the control population. Genetic mutations have been reported as potential contributing factors to pathogenesis of glaucoma. In this study, we reported a Chinese family that many members affected with primary open‐angle glaucoma carried a PAPPA2 c.392G>C heterozygous mutation. Western blotting results showed that this mutation decreased PAPPA2 protein level. Strikingly, we found PAPPA2 and its substrate IGFBP5 both expressed in human aqueous humor samples, and PAPPA2 levels significantly decreased in the POAG group accompanied with IGFBP5 levels increased in the POAG group. We also demonstrated that PAPPA2 cleaved IGFBP5 in vitro. We found that overexpressed IGFBP5 increased many fibrosis‐related gene expression through mRNA sequencing, western blotting, and immunofluorescence staining assays in primary human trabecular meshwork cells (HTMCs). More importantly, we found an inadequate dose of Pappa2 caused POAG‐like phenotypes in the mouse model. So, we proved the vital function of the eye local PAPPA2‐IGFBP5 axis in regulating extracellular matrix homeostasis and contributing to trabecular meshwork fibrosis and pathogenesis of POAG.