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gelatine

3D hydrogel-

based cell culture

gel preparation

Sources & composition

NATURAL

+ cell adhesion sites

+ cell-mediated degradation

- batch-to-batch variation

- limited range of properties

- sterility / endotoxins

SYNTHETIC

+ tailorable

+ reproducible

+ GMP

- lack of biological

functionality

- expensive

Klotz, ..., Melchels 2016 Trends in Biotechnology

polyacrylamide

PEG-DMA

PEG-PLGA

pHPMAm-

lactate-PEG

DNA-polypeptide

poloxamer-DA

proteins

collagen

gelatin

fibrin

elastin

silk

polysaccharides

alginate

aggarose

methylcellulose

carrageenan

chitosan

hyaluronic acid

HYBRID

gelMA/gellan

HA-SH + PEG-A

HA + dex-HEMA

PEG-DMA + gelMA

PEG-DA + alginate

Benya PD, Shaffer JD. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels.

Cell 1982;30:215–24

Dunn JCY, Yarmush ML, Koebe HG, Tompkins RG. Hepatocyte function and extracellular matrix geometry long-term culture in a sandwich configuration.

FASEB J 1989;3:174–7.

dECM

fat

cartilage

heart

Schuurman & Levett, ..., Melchels et al 2013 Macromolecular Bioscience

gelMA for

3D cell culture

What is a hydrogel?

cancer

Hydrogels

OV-MZ-6 ovary cancer cell line

Crosslinks can be chemical or physical

cartilage

Single-component hydrogels

Applications

Multi-component hydrogels

Levett, Melchels et al 2013 J Biomed Mater Res A

fused deposition modelling (FDM)

SLS/SLM/EBM

Acknowledgements

(projection)

stereolithography (SLA)

hydrogels are highly suited artificial extracellular matrices

3D biofabricaton techniques be used to provide spatial organisation

to cells and biomaterials

Fabrication

techniques

Lim, ..., Melchels et al 2018 Biofabrication

Hydrogel-based 3D Biofabrication

Strategies

combining biofabrication

with cellular self-organisation

provides a potent strategy for

engineering complex tissues

Dr Ferry PW Melchels

jet-based printing

Scaffolding

Pre-fabricated scaffolds

Hochleitner et al 2015 Biofabrication

extrusion

Scaffold materials for BIOprinting

Melchels & Blokzijl et al. 2016 Biofabrication

PCL, PCL and ..... PCL

Cell-supporting

(bio-inks)

gelMA/gellan

Bio-inks

1) yielding

Material

functions

2) shear-thinning

gelatine-methacryloyl (gelMA)

Actuation

Facilitate fabrication

Create vasculature

Printed removable moulds

MACRO

MICRO + MACRO

Rheology modifier

Suspension baths

Sensing

Hinton et al. 2015 Science Advances

Bhattacharjee et al. 2015 Science Advances

Keriquel et al 2010 Biofabrication

Mouser, Melchels et al 2016 Biofabrication

Gladman et al. 2016 Nature Materials

Chang et al. 2011 J Biomed Mater Res B Appl Biomater.

metals

Ti

CoCrMo

Ti6Al4V

316SS

Xu et al 2009 Biofabrication

Visser, Melchels et al. 2015 Nature Communications

Kämmerer, Melchels et al 2014 Acta Biomaterialia

hydrogels

PEG-DA + alginate + nanoclay

Gellan gum:

  • polysaccharide
  • degradable
  • high MW -> high viscosity
  • ionic crosslinking
  • chondrogenic

Melchels et al. 2014 Journal of Materials Chemistry part B

Dietmar Hutmacher

QUT

Dirk Grijpma

UTwente

Jos Malda

UMC Utrecht

Mannoor et al. 2013 Nano Letters

PETA.org

blue = 20M cells/mL

red = 60M cells/mL

Luo et al, 2013 J Mater Chem B

Kämmerer, Melchels et al 2014 Acta Biomaterialia

bioactive glass

Melchels & Blokzijl et al. 2016 Biofabrication

Paxton, ..., Melchels et al. 2017 Biofabrication

* denatured extracellular matrix

* cell adhesion sites

* enzymatically degradable

* non-immunogenic

* cheap and widely available

* thermosensitive

thermosets

PCL

PDLLA

PTMC

PPF

Schuurman et al. 2011 Biofabrication

Caliari et al 2016 Nature Methods

Pati et al. 2014 Nature Communications

ceramics

TCP

HAP

CaSiO3

Al2O3

SiO2

Mg3(PO4)2

Kesti & Müller et al. 2015 Acta Biomaterialia

MICRO

endothelial cells in Matrigel

making a capillary network

Kubota et al 1988 Journal of Cell Biology

Klotz, ..., Melchels et al European Cells and Materials (submitted)

graphene

Fluorescence Recovery After Photo-bleaching (FRAP)

Loessner, ..., Melchels et al 2016 Nature Protocols

collagen II aggrecan collagen I

explainingthefuture.com

Spiller et al 2011 Tissue Eng Part B Rev

Melchels et al. 2014 J Mater Chem B

Kolesky et al. 2014 Advanced Materials + 2016 PNAS

thermoplastics

PL(G)A

PCL

PTMC

PolyActive

PHB(V)

chitosan

gelMA PEG-MA

HA-MA AL-MA

54x

Malda et al. 2013 Advanced Materials

3) photo-curing

Levett, Melchels et al 2014 Acta Biomaterialia

Visser, Melchels et al. 2015 Nature Communications

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