Mouse in-vitro spermatogenesis on alginate-based 3D bioprinted constructs

Commonly used acronym: IVS

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Scope of the method

The Method relates to

Animal health
Human health

The Method is situated in

Basic Research

Type of method

In chemico

Species from which cells/tissues/organs are derived

Mouse

Type of cells/tissues/organs

Testis

Description

Method keywords

3D in vitro model
3D bioprinting
organoid culture
microscaffolds
extrusion-based 3D printing

Scientific area keywords

fertility preservation
reprotoxicity
preclinical
Drug discovery
drug development
testicular physiology

Method description

Studying spermatogenesis in situ has led to the understanding that the 3D reorganization of testicular cells into an interstitial and tubular compartment is of enormous importance for germ cell differentiation. We will rely on 3D bioprinting technology which gives control over cell deposition and scaffold design, to recreate the compartmentalization of the testis in vitro. Testicular constructs will be produced by culturing epithelial testicular cell fractions in the macropores of bioprinted interstitial cell-laden scaffolds. We expect these biomimetic scaffolds will also support differentiation of human germ cells.

Lab equipment

Extrusion-based 3D printer;
Air compressor;
Hydrogel;
Fluorescence activated cell sorter or magnetic activated cell sorter;
Fluorescence microscope.

Method status

Still in development
Published in peer reviewed journal

Pros, cons & Future potential

Advantages

Allows manipulation of cell suspensions before culture to help understand the many mechanisms controlling testicular physiology and spermatogenesis, but also to discover new clinical targets.

Challenges

Requires high cell concentrations ;
Lack of bioactivity or biocompatibility of the hydrogel ;
Uncertainty related to the medium ingredients that drive testicular morphogenesis and spermatogenesis.

Modifications

Use of higher cell concentrations ;
Use of alternative hydrogels ;
Optimisation culture medium.

Future & Other applications

Tool to study testicular physiology through cell manipulation or gene editing ;
In vitro derived sperm of prepubertal cancer patients and adult non-obstructive patients can be used to generate offspring through assisted reproduction ;
Cell therapy ;
Following the incorporation into multi-organs microfluidic devices, the constructs can serve as a high-throughput screening assay in preclinical tests.

References, associated documents and other information

References

Baert, Y., Dvorakova-Hortova, K., Margaryan, H. & Goossens, E. (2019) Mouse in vitro spermatogenesis on alginate-based bioprinted scaffolds. Biofabrication 11, 035011

Richer, G., Hobbs, R., Loveland, Kate., Goossens, E., Baert, Y. (2021) Long-term maintenance and meiotic entry of early germ cells in murine testicular organoids functionalized by 3D printed scaffolds at the air-medium interface cultivation. Frontiers in Physiology 12, 757565

Associated documents

Baert_2019_Biofabrication.pdf
Richer et al. 2021.pdf