This document discusses Matrigel and its applications in tissue engineering. Some key points: - Matrigel is a natural hydrogel extracted from mouse tumors that mimics the extracellular matrix. It promotes cell differentiation and adhesion. - While it provides an excellent environment for cells, Matrigel has disadvantages like uncontrolled composition and poor mechanical properties. - 3D bioprinting allows the precise placement of multiple cell types and materials within hydrogel structures. Studies have used Matrigel to print vascularized bone and contractile cardiac tissues. - Matrigel is widely used in regenerative medicine as a scaffold to deliver cells and growth factors. It has applications in skin grafts, cancer models,

1. Matrigel andMatrigel and
Its ApplicationsIts Applications
Olga Filippova, Akash Patel

2. – Temperature Control System
– Cool Matrigel Reservoir
– Heat substrate
– Material Delivery System
– Motion System
Temperature Control System for Solid Freeform
Fabrication of Cell-Laden Matrigel Constructs
ProposalProposal

3. – Lobule = Functional Unit
– Hepatocytes
– 70 – 80% of liver mass
– Sinusoidal plates
– Major liver functions
– Drug metabolism
– ECM
– Collagens
– I, III, IV, V, XVIII
– Glycoproteins
– Laminin, Fibronectin
Liver IntroductionLiver Introduction
www.ece.ncsu.edu/imaging/MedImg/SIMS/GE1_3.html

4. – Natural hydrogel
– Basement membrane from EHS mouse sarcoma
– Advantages
– Composition
– Laminin
– Collagen IV
– Thermal Crosslinking
– 4o
C (liquid)  37o
C (gel)
– Outstanding biocompatibility
– Promotes differentiation
Gelain et al
MatrigelMatrigel

5. – Disadvantages
– Uncontrolled composition
– Poor gelation kinetics
– Little cell proliferation
– Lower mechanical integrity
– Young’s Modulus 400 Pa – highly compliable
– Shear Modulus 180 Pa – below natural liver tissue
– Thermal Crosslinking
– Reverse characteristics – liquid at low temperatures, solid at
physiological temperatures
Matrigel (cont)Matrigel (cont)

6. – Visualization
– Proper sinusoidal shape
– Proper width
– Gelation
– Time
– Mass/Volume loss
– Mechanical Testing
– Compression
Construct CharacterizationConstruct Characterization

7. – Cell Viability
– Live/Dead Assay – membrane damage
– Cell Proliferation
– Alamar Blue Assay – growth environment analysis
– Cell-Specific Function
– Quanti Chrome Assay – urea production
Cell CharacterizationCell Characterization
http://www.ab-direct.com/about/alamarblue-483.htmlHuuskonen et al

8. – Especially useful in regenerative medicine
– May help solve the problem of limited donor grafts
– Scaffold optimization with tailored biological properties
– Provide:
– Supportive environment for cell attachment, proliferation, and differentiation
– Biological cues to elicit desired cellular response
– ECM composition, growth factors
Great Promise for 3D Computer-AidedGreat Promise for 3D Computer-Aided
Deposition of Cell-Laden Hydrogel StructureDeposition of Cell-Laden Hydrogel Structure

9. – 3D printed cell laden spatially organized hydrogel scaffolds
– Demonstrate ability to print two distinct cell populations at predefined locations
– Osteogenic and endothelial progenitors
– Potentially use 3D printing to develop vascularized bone grafts for TE
* Air humidity during printing process may influence survival of extruded cells
5 hrs
Day 1
Day 2
Day 3
Matrigel Lutrol E127 25%
viablecells(%oftotal)
Alginate 2% Agarose 1%
Bone Tissue PrintingBone Tissue Printing

10. Bone Tissue PrintingBone Tissue Printing

11. – Simultaneous printing of cells and biomaterials allows precise placement of cells and
proteins within 3D hydrogel structures
– Created contractile cardiac hybrids have properties that can be tailored in 3D to achieve
desired porosities, mechanical and chemical properties
– Results suggest that the printing method could be used for hierarchical design of functional
cardiac patches, balanced with porosity for mass transport and structural support
Drop-on-Demand Printing of Cell andDrop-on-Demand Printing of Cell and
Materials for Designer Tissue ConstructsMaterials for Designer Tissue Constructs

12. – Use of synthetic degradable gels is emerging
–Matrigel
– Primarily as a way to deliver cells and/or molecules
– In situ
– Smart matrices
– Highly specific 3D architecture for skin grafts
Tissue EngineeringTissue Engineering
Replacement SkinReplacement Skin

13. – 3D cell cultures have much greater potential than 2D cultures
– Using 3D cultures, scientists discover patterns of gene
expression and other biological activities that more closely
mirror what happens in organisms
– 3D ECM models are especially useful for monitoring
mechanisms of tumor growth and metastasis
Cancer Research ModelsCancer Research Models

14. “3-D microwell culture of human embryonic stem cells”
– Constructed a 3-D microwell
system for long-term hESC culture
– Physical and extracellular matrix
patterning constraints limit colony
growth
– Matrigel used to culture cells
Other Demands for MicroscaleOther Demands for Microscale
Accurate, Precise Matrigel PrintingAccurate, Precise Matrigel Printing

15. Growth of miniature pig parotid cells on biomaterials in vitro
– Coating the surface of synthetic materials aids cell growth and maintenance of a
morphology that more closely resembles normal epithelium
Compatibility of human fetal neural stem cells with hydrogel biomaterials
in vitro
– Stroke and spinal cord or brain injury often result in cavity formation
– Stem cell transplantation in combination with tissue engineering has the potential to
fill cavity and replace lost neurons
– Hydrogels (Matrigel) may enhance migration capability across the injured cavity in
vivo due to their ability to provide a three-dimensional matrix scaffold suitable for
cellular adhesion, migration and support
Other Uses for MatrigelOther Uses for Matrigel
Printing DevicePrinting Device

16. Craniofacial muscle engineering using a 3-dimensional phosphate glass
fibre construct
– A 3-dimensional mesh arrangement of the glass fibres was the best at
encouraging cell attachment and proliferation
– In addition, increasing the density of the seeded cells and using Matrigel and
insulin-like growth factor I enhanced the formation of prototypic muscle fibers
Beyond the Cells: Scaffold Matrix Character Affects the In Vivo
Performance of Purified Adipocyte Fat Grafts
– Focused on the effects of fat graft structure on in vivo performance
– Adult adipocyte mixed with Matrigel were implanted as fat grafts
– Purified Matrigel grafts showed statistically greater longevity and volume
maintenance versus all other groups
Other Uses for MatrigelOther Uses for Matrigel
Printing Device (cont)Printing Device (cont)