期刊
ADVANCED HEALTHCARE MATERIALS
卷 11, 期 21, 页码 -出版社
WILEY
DOI: 10.1002/adhm.202201826
关键词
3D model systems; astrocytes; Ca2+-Imaging; cortical neurons; hyaluronic acid; melt electrowriting
资金
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [326998133 - TRR 225]
- GSLS Wuerzburg, Germany
- Projekt DEAL
3D neuronal cultures using an ultra-soft matrix reinforced with a microfiber frame show improved neuronal viability and network formation. By pre-seeding astrocytes, the neuronal survival and firing activity can be further enhanced, making this system a reliable and reproducible tool for studying brain diseases.
3D neuronal cultures attempt to better replicate the in vivo environment to study neurological/neurodegenerative diseases compared to 2D models. A challenge to establish 3D neuron culture models is the low elastic modulus (30-500 Pa) of the native brain. Here, an ultra-soft matrix based on thiolated hyaluronic acid (HA-SH) reinforced with a microfiber frame is formulated and used. Hyaluronic acid represents an essential component of the brain extracellular matrix (ECM). Box-shaped frames with a microfiber spacing of 200 mu m composed of 10-layers of poly(e-caprolactone) (PCL) microfibers (9.7 +/- 0.2 mu m) made via melt electrowriting (MEW) are used to reinforce the HA-SH matrix which has an elastic modulus of 95 Pa. The neuronal viability is low in pure HA-SH matrix, however, when astrocytes are pre-seeded below this reinforced construct, they significantly support neuronal survival, network formation quantified by neurite length, and neuronal firing shown by Ca2+ imaging. The astrocyte-seeded HA-SH matrix is able to match the neuronal viability to the level of Matrigel, a gold standard matrix for neuronal culture for over two decades. Thus, this 3D MEW frame reinforced HA-SH composite with neurons and astrocytes constitutes a reliable and reproducible system to further study brain diseases.
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