Biofabrication and Characterization of Vascularizing PEG-Norbornene Microgels

Authors:
Nicole E Friend , Irene W Zhang , Michael M Hu , Atticus J McCoy , Robert N Kent 3rd Samuel J DePalma , Brendon M Baker , Sasha Cai Lesher-Pérez , Jan P Stegemann , Andrew J Putnam 
In:
Source: Biofabrication
Publication Date: (2025)
Issue: 113: 4
Research Area:
Regenerative medicine
Cells used in publication:
Endothelial, umbilical vein, human (HUVEC)
Species: human
Tissue Origin: vein
Fibroblast, lung, human normal (NHLF)
Species: human
Tissue Origin: lung
Experiment

2.1 Cell Culture

Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical cords from the University of Michigan Mott Children's Hospital as previously described [39]. Umbilical cords were obtained by a process considered exempt by the University of Michigan's Institutional Review Board (notice of determination dated August 21, 2014) because the tissue is normally discarded, and no identifying information is provided to the researchers who receive the cords. HUVEC were cultured in fully supplemented EGM2 (Lonza Inc., Walkersville, MD). HUVEC were used from passages 4 to 7. Normal human lung fibroblasts (NHLFs; Lonza) were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco, Waltham, MA) supplemented with 10% fetal bovine serum (FBS; Gibco). NHLF were used from passages 10 to 15. All cells were cultured at 37°C and 5% CO2 with media replacement every 2?days.

Abstract

Establishing a robust, functional microvascular network remains a critical challenge for both the revascularization of damaged or diseased tissues and the development of engineered biological materials. Vascularizing microgels may aid in efforts to develop complex, multiphasic tissues by providing discrete, vascularized tissue modules that can be distributed throughout engineered constructs to vascularize large volumes. Here, we fabricated poly(ethylene glycol)-norbornene (PEGNB) microgels containing endothelial and stromal cells via flow-focusing microfluidic droplet generation. When embedded in bulk fibrin hydrogels, these cell-laden microgels initiated the formation and development of robust microvascular networks. Furthermore, extended preculture of cell-laden PEGNB microgels enabled the formation of vessel-like structures supported by basement membrane within the matrix without aggregation. Our findings highlight the suitability of PEG-based matrices for the development of vascularizing microgels capable of forming well-distributed, robust microvascular networks.