Matrigel and Matrigel and Its ApplicationsIts Applications

Olga Filippova, Akash Patel

– 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

– 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

– Natural hydrogel– Basement membrane from EHS mouse sarcoma

– Advantages– Composition

– Laminin

– Collagen IV

– Thermal Crosslinking – 4oC (liquid) 37oC (gel)

– Outstanding biocompatibility

– Promotes differentiation

Gelain et al

MatrigelMatrigel

– 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)

– Visualization– Proper sinusoidal shape– Proper width

– Gelation– Time– Mass/Volume loss

– Mechanical Testing– Compression

Construct CharacterizationConstruct Characterization

– 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

– 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-Aided Great Promise for 3D Computer-Aided Deposition of Cell-Laden Hydrogel Structure Deposition of Cell-Laden Hydrogel Structure

– 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

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Day 2

Day 3

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Alginate 2% Agarose 1%

Bone Tissue PrintingBone Tissue Printing

Bone Tissue PrintingBone Tissue Printing

– 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 and Drop-on-Demand Printing of Cell and Materials for Designer Tissue ConstructsMaterials for Designer Tissue Constructs

– 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 Engineering Tissue Engineering Replacement SkinReplacement Skin

– 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

“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

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 Matrigel Other Uses for Matrigel Printing DevicePrinting Device

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 Matrigel Other Uses for Matrigel Printing Device (cont)Printing Device (cont)