Bio-printing cell-laden Matrigel–agarose constructs

3D printing of biological architectures that mimic the structural and functional features of in vivo tissues is of great interest in tissue engineering and the development of transplantable organ constructs. Printable bio-inks that are compatible with cellular activities play critical roles in the process of 3D bio-printing. Although a variety of hydrogels have been used as bio-inks for 3D bio-printing, they inherit poor mechanical properties and/or the lack of essential protein components that compromise their performance. Here, a hybrid Matrigel–agarose hydrogel system has been demonstrated that possesses both desired rheological properties for bio-printing and biocompatibility for long-term (11 days) cell culture. The agarose component in the hybrid hydrogel system enables the maintenance of 3D-printed structures, whereas Matrigel provides essential microenvironments for cell growth. When human intestinal epithelial HCT116 cells are encapsulated in the printed Matrigel–agarose constructs, high cell viability and proper cell spreading morphology are observed. Given that Matrigel is used extensively for 3D cell culturing, the developed 3D-printable Matrigel–agarose system will open a new way to construct Matrigel-based 3D constructs for cell culture and tissue engineering.

[1]  Wouter J A Dhert,et al.  Distinct tissue formation by heterogeneous printing of osteo- and endothelial progenitor cells. , 2011, Tissue engineering. Part A.

[2]  L. Niklason,et al.  Scaffold-free vascular tissue engineering using bioprinting. , 2009, Biomaterials.

[3]  Fabrizio Gelain,et al.  Designer Self-Assembling Peptide Nanofiber Scaffolds for Adult Mouse Neural Stem Cell 3-Dimensional Cultures , 2006, PloS one.

[4]  Mark W. Tibbitt,et al.  Hydrogels as extracellular matrix mimics for 3D cell culture. , 2009, Biotechnology and bioengineering.

[5]  S. Rizzi,et al.  Elucidating the role of matrix stiffness in 3D cell migration and remodeling. , 2011, Biophysical journal.

[6]  Brendon M. Baker,et al.  Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues , 2012 .

[7]  B. Duan,et al.  3D bioprinting of heterogeneous aortic valve conduits with alginate/gelatin hydrogels. , 2013, Journal of biomedical materials research. Part A.

[8]  Xiang-Yang Liu,et al.  Topology evolution and gelation mechanism of agarose gel. , 2005, The journal of physical chemistry. B.

[9]  Takao Hayakawa,et al.  3D spheroid culture of hESC/hiPSC-derived hepatocyte-like cells for drug toxicity testing. , 2013, Biomaterials.

[10]  R. Wells The role of matrix stiffness in regulating cell behavior , 2008, Hepatology.

[11]  Wei Sun,et al.  Bioprinting endothelial cells with alginate for 3D tissue constructs. , 2009, Journal of biomechanical engineering.

[12]  Jun Liu,et al.  Monitoring nutrient transport in tissue‐engineered grafts , 2015, Journal of tissue engineering and regenerative medicine.

[13]  P. Vogt,et al.  Tissue Engineered Skin Substitutes Created by Laser-Assisted Bioprinting Form Skin-Like Structures in the Dorsal Skin Fold Chamber in Mice , 2013, PloS one.

[14]  David A Stenger,et al.  Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices , 2001, Neuroscience Letters.

[15]  B. Radotra,et al.  GLIOMA INVASION IN VITRO IS MEDIATED BY CD44–HYALURONAN INTERACTIONS , 1997, The Journal of pathology.

[16]  Matthias P. Lutolf,et al.  Designing materials to direct stem-cell fate , 2009, Nature.

[17]  Hon Fai Chan,et al.  3D Printing of Highly Stretchable and Tough Hydrogels into Complex, Cellularized Structures , 2015, Advanced materials.

[18]  Yinglin Xia,et al.  Salmonella‐infected crypt‐derived intestinal organoid culture system for host–bacterial interactions , 2014, Physiological reports.

[19]  Farshid Guilak,et al.  Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds. , 2004, Biomaterials.

[20]  Ying Luo,et al.  A photolabile hydrogel for guided three-dimensional cell growth and migration , 2004, Nature materials.

[21]  Ying Mei,et al.  3D printing facilitated scaffold-free tissue unit fabrication , 2014, Biofabrication.

[22]  H. Clevers,et al.  Growing Self-Organizing Mini-Guts from a Single Intestinal Stem Cell: Mechanism and Applications , 2013, Science.

[23]  H. Fischer,et al.  Three-dimensional printing of stem cell-laden hydrogels submerged in a hydrophobic high-density fluid , 2012, Biofabrication.

[24]  R. M. Sharrard,et al.  Prostate epithelial cell lines form spheroids with evidence of glandular differentiation in three-dimensional Matrigel cultures , 2001, British Journal of Cancer.

[25]  Zhongmin Jin,et al.  Sequential assembly of 3D perfusable microfluidic hydrogels , 2014, Journal of Materials Science: Materials in Medicine.

[26]  D. D’Lima,et al.  Direct human cartilage repair using three-dimensional bioprinting technology. , 2012, Tissue engineering. Part A.

[27]  Nupura S. Bhise,et al.  Direct-write bioprinting of cell-laden methacrylated gelatin hydrogels , 2014, Biofabrication.

[28]  Jackie Y Ying,et al.  The effect of matrix stiffness on mesenchymal stem cell differentiation in a 3D thixotropic gel. , 2010, Biomaterials.

[29]  David J. Mooney,et al.  Harnessing Traction-Mediated Manipulation of the Cell-Matrix Interface to Control Stem Cell Fate , 2010, Nature materials.

[30]  Beum Jun Kim,et al.  Designing compartmentalized hydrogel microparticles for cell encapsulation and scalable 3D cell culture. , 2015, Journal of materials chemistry. B.

[31]  Rong Fan,et al.  Leaf-inspired artificial microvascular networks (LIAMN) for three-dimensional cell culture , 2015 .

[32]  James J. Yoo,et al.  Bioprinted Amniotic Fluid‐Derived Stem Cells Accelerate Healing of Large Skin Wounds , 2012, Stem cells translational medicine.

[33]  T. Hasan,et al.  A three-dimensional in vitro ovarian cancer coculture model using a high-throughput cell patterning platform. , 2011, Biotechnology journal.

[34]  R V Bellamkonda,et al.  Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. , 2001, Biomaterials.

[35]  Nancy L Allbritton,et al.  Optimization of 3-D organotypic primary colonic cultures for organ-on-chip applications , 2014, Journal of Biological Engineering.

[36]  Glenn D Prestwich,et al.  Bioprinting vessel-like constructs using hyaluronan hydrogels crosslinked with tetrahedral polyethylene glycol tetracrylates. , 2010, Biomaterials.