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Perfusable Vascular Network with a Tissue Model in a Microfluidic Device
Published on: April 4, 2018
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A robust vasculogenic microfluidic model using human immortalized endothelial cells and Thy1 positive fibroblasts
Zhengpeng Wan1, Shun Zhang2, Amy X Zhong2
1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
Biomaterials
|July 24, 2021
Summary
We developed stable cell lines for creating microvascular networks in vitro. Fibroblast Thy1 expression is crucial for functional network formation, with later passage cells showing reduced function.
Area of Science:
- * Regenerative Medicine
- * Cell Biology
- * Tissue Engineering
Background:
- * Primary human umbilical vein endothelial cells (HUVECs) and fibroblasts (FBs) are used for in vitro microvascular network (μVN) formation.
- * Donor variability and limited lifespan of primary cells hinder reproducible μVN generation.
- * Immortalization of HUVECs and FBs using human telomerase reverse transcriptase (hTERT) offers a solution.
Purpose of the Study:
- * To establish stable cell sources for reproducible in vitro μVN formation.
- * To investigate the role of fibroblast Thy1 expression in μVN development and function.
- * To identify methods for maintaining functional fibroblast characteristics for improved μVN perfusion.
Main Methods:
- * Immortalization of HUVECs and FBs by introducing hTERT.
- * Co-culture of immortalized cells to form μVNs.
- * Analysis of Thy1 expression in fibroblasts across different passages.
- * Assessment of fibroblast-secreted factors (IGFBP2, IGFBP7, SPARC) and their role in angiogenesis.
- * Evaluation of μVN perfusability and microvessel obstruction.
- * Optimization of fibroblast culture conditions (gelatin coating, serum-free medium).
Main Results:
- * Immortalized HUVECs and FBs can form μVNs, but function depends on fibroblast passage number.
- * Thy1 expression decreases in later passage fibroblasts.
- * Thy1-positive fibroblasts exhibit higher expression of IGFBP2, IGFBP7, and SPARC, promoting angiogenesis and lumen formation.
- * Thy1-negative fibroblasts impede microvessel openings, reducing μVN perfusability.
- * Culturing immortalized FBs on gelatin in serum-free medium preserves Thy1-positive cells and supports perfusable μVN formation.
Conclusions:
- * Immortalized cell lines provide stable sources for μVN generation.
- * Fibroblast Thy1 expression is a critical regulator of in vitro vasculogenesis and μVN function.
- * Maintaining Thy1-positive fibroblast populations is essential for creating perfusable microvascular networks.
