Photopatterning of hydrogel scaffolds coupled to filter materials using stereolithography for perfused 3D culture of hepatocytes.
In vitro models that recapitulate the liver's structural and functional complexity could prolong hepatocellular viability and function to improve platforms for drug toxicity studies and understanding liver pathophysiology. Here, stereolithography (SLA) was employed to fabricate hydrogel scaffolds with open channels designed for post-seeding and perfused culture of primary hepatocytes that form 3D structures in a bioreactor. Photopolymerizable polyethylene glycol-based hydrogels were fabricated coupled to chemically activated, commercially available filters (polycarbonate and polyvinylidene fluoride) using a chemistry that permitted cell viability, and was robust enough to withstand perfused culture of up to 1 µL/s for at least 7 days. SLA energy dose, photoinitiator concentrations, and pretreatment conditions were screened to determine conditions that maximized cell viability and hydrogel bonding to the filter. Multiple open channel geometries were readily achieved, and included ellipses and rectangles. Rectangular open channels employed for subsequent studies had final dimensions on the order of 350 µm by 850 µm. Cell seeding densities and flow rates that promoted cell viability were determined. Perfused culture of primary hepatocytes in hydrogel scaffolds in the presence of soluble epidermal growth factor (EGF) prolonged the maintenance of albumin production throughout the 7-day culture relative to 2D controls. This technique of bonding hydrogel scaffolds can be employed to fabricate soft scaffolds for a number of bioreactor configurations and applications.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139.
Jaclyn A Shepard
JA
Neiman
Ritu
R
Raman
Vincent
V
Chan
Mary G
MG
Rhoads
Micha Sam B
MS
Raredon
Jeremy J
JJ
Velazquez
Rachel L
RL
Dyer
Rashid
R
Bashir
Paula T
PT
Hammond
Linda G
LG
Griffith
D019149:Bioreactors; D002470:Cell Survival; D004815:Epidermal Growth Factor / Q000378:metabolism; D022781:Hepatocytes / Q000502:physiology*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D009924:Organ Culture Techniques / Q000379:methods*; D054457:Tissue Scaffolds
D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D004815:Epidermal Growth Factor
(3-6): hydrogel; hepatocyte; liver; patterning; stereolithography