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Enhancing maturity in 3D kidney micro-tissues through clonogenic cell combinations and endothelial integration
Fatemeh Abdollahzadeh1,2, Niloofar Khoshdel-Rad1, Khadijeh Bahrehbar1
1Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Journal of Cellular and Molecular Medicine
|May 31, 2024
Summary
Researchers developed a simple method to create advanced 3D kidney organoids from mouse renal cells. These organoids, when combined with endothelial cells, show enhanced maturity and gene expression, paving the way for better kidney disease research.
Area of Science:
- Biomedical Engineering
- Developmental Biology
- Regenerative Medicine
Background:
- Three-dimensional (3D) organoid culture is an advanced laboratory model for studying kidney development, physiology, and abnormalities.
- Kidney organoids derived from primary renal cells mimic key characteristics of native kidney tissue.
- Current methods for generating complex kidney organoids require optimization for improved maturity and functionality.
Purpose of the Study:
- To present a simple method for isolating and characterizing mouse renal clonogenic mesenchymal (MLCs) and epithelial-like cells (ELCs).
- To generate mature 3D kidney micro-tissues (MEHRMs) using isolated MLCs and ELCs, with and without human umbilical vein endothelial cells (HUVECs).
- To evaluate the maturity and specific gene expression of the generated 3D kidney micro-tissues.
Main Methods:
- Isolation and characterization of mouse renal clonogenic mesenchymal (MLCs) and epithelial-like cells (ELCs) using flow cytometry.
- Culture of MLCs and ELCs alone (ME) or in combination with HUVECs (MEH) at a 10:5:2 ratio.
- Analysis of 3D MEH renal micro-tissues (MEHRMs) for renal-specific gene expression and ex ovo transplantation for maturation assessment.
Main Results:
- MLCs expressed key surface markers (Cd44, Cd73, Cd105) in over 93% of cells.
- ELCs showed upregulated epithelial and stem/progenitor cell markers.
- 3D MEH renal micro-tissues exhibited significantly increased expression of renal-specific genes (Aqp1, Cdh1, Umod, Wt1, Podxl, Nphs1) compared to tissues without endothelial cells.
- Ex ovo transplantation demonstrated enhanced maturation of the constructed 3D kidney.
Conclusions:
- The presented method effectively isolates and characterizes mouse renal mesenchymal and epithelial cells.
- Co-culture with endothelial cells significantly enhances the maturity and specific gene expression of 3D kidney organoids.
- These advanced 3D kidney organoids represent a promising model for studying kidney development and disease.
Keywords:
clonogenic epithelial‐like cellsclonogenic mesenchymal‐like cellsco‐culturekidney micro‐tissues
