scholarly journals Recombinant Spider Silk Protein Matrices Facilitate Differentiation of Neural Stem Cells Into Mature and Functional Neurons

2021 ◽  
Vol 7 ◽  
Author(s):  
Michalina Lewicka ◽  
Paola Rebellato ◽  
Jakub Lewicki ◽  
Per Uhlén ◽  
Anna Rising ◽  
...  

Neural stem cells (NSCs) show great promise in drug discovery and clinical application. Yet few efforts have been made to optimize biocompatible materials for such cells to be expanded and used in clinical conditions. We have previously demonstrated that NSCs are readily cultured on substrates of certain recombinant spider silk protein without addition of animal- or human-derived components. The question remains however whether this material allows differentiation into functional neurons, and whether such differentiation can take place also when the NSCs are cultured not only upon but also within the biodegradable material. Here we demonstrate that “foam”-like structures generated from recombinant spider silk protein (4RepCT) provided excellent matrices for the generation and multicellular analysis of functional excitatory neurons from NSCs without addition of animal- or human-derived components. NSCs isolated from the cerebral cortices of rat embryos were cultured at either 4RepCT matrices shaped as foam-like structures without coating, or on conventional polystyrene plates coated with poly-L-ornithine and fibronectin. Upon treatment with recombinant proteins including the extracellular signaling factor BMP4 or a combination of BMP4 and the signaling factor Wnt3a, the cortical NSCs cultured in 4RepCT foam-like structures differentiated efficiently into neurons that responded to glutamate receptor agonists, such as AMPA, to the same extent as control cultures. Matrices derived from recombinant spider silk proteins thus provide a functional microenvironment for neural stem cells with little or no animal- or human-derived components and can be employed in the development of new strategies in stem cell research and tissue engineering.

2019 ◽  
Author(s):  
Michalina Lewicka ◽  
Paola Rebellato ◽  
Jakub Lewicki ◽  
Per Uhlén ◽  
Anna Rising ◽  
...  

Neural progenitors or stem cells (NSCs) show great promise in drug discovery and clinical application. Yet few efforts have been made to optimize biocompatible materials for such cells to be expanded and used in clinical conditions. We have previously demonstrated that NSCs are readily cultured on substrates of certain recombinant spider silk protein without addition of animal- or human-derived components. The question remains however whether this material allows differentiation into functional neurons and glia, and whether such differentiation can take place also when the NSCs are cultured within the material in a pseudo-3D context. Here we demonstrate that “foam”-like structures generated from recombinant spider silk protein (4RepCT) provided excellent matrices for the generation and multicellular analysis of functional excitatory neurons from NSCs without addition of animal- or human-derived components. NSCs isolated from the cerebral cortices of rat embryos were cultured on either 4RepCT matrices shaped as foam-like structures without coating, or on conventional polystyrene plates coated with poly-L-ornithine and fibronectin. Upon treatment with recombinant proteins including the growth factor BMP4 or a combination of BMP4 and the signaling factor Wnt3a, the cortical NSCs cultured in 4RepCT foam-like structures differentiated efficiently into neurons that responded to glutamate receptor agonists, such as AMPA, to at least the same extent as control cultures. Matrices derived from recombinant spider silk proteins thus provide a functional microenvironment for neural stem cells without any animal- or human-derived components, and can be employed in the development of new strategies in stem cell research and tissue engineering.


Molecules ◽  
2022 ◽  
Vol 27 (2) ◽  
pp. 511
Author(s):  
Yu Suzuki ◽  
Takanori Higashi ◽  
Takahiro Yamamoto ◽  
Hideyasu Okamura ◽  
Takehiro K. Sato ◽  
...  

Spider dragline silk is a biopolymer with excellent mechanical properties. The development of recombinant spider silk protein (RSP)-based materials with these properties is desirable. Formic acid (FA) is a spinning solvent for regenerated Bombyx mori silk fiber with excellent mechanical properties. To use FA as a spinning solvent for RSP with the sequence of major ampullate spider silk protein from Araneus diadematus, we determined the conformation of RSP in FA using solution NMR to determine the role of FA as a spinning solvent. We assigned 1H, 13C, and 15N chemical shifts to 32-residue repetitive sequences, including polyAla and Gly-rich regions of RSP. Chemical shift evaluation revealed that RSP is in mainly random coil conformation with partially type II β-turn structure in the Gly-Pro-Gly-X motifs of the Gly-rich region in FA, which was confirmed by the 15N NOE data. In addition, formylation at the Ser OH groups occurred in FA. Furthermore, we evaluated the conformation of the as-cast film of RSP dissolved in FA using solid-state NMR and found that β-sheet structure was predominantly formed.


Langmuir ◽  
2018 ◽  
Vol 34 (39) ◽  
pp. 11795-11805 ◽  
Author(s):  
Linnea Nilebäck ◽  
Suvi Arola ◽  
Mathias Kvick ◽  
Arja Paananen ◽  
Markus B. Linder ◽  
...  

2014 ◽  
Vol 14 (7) ◽  
pp. 936-942 ◽  
Author(s):  
John G. Hardy ◽  
André Pfaff ◽  
Aldo Leal-Egaña ◽  
Axel H. E. Müller ◽  
Thomas R. Scheibel

2011 ◽  
Vol 14 (3) ◽  
pp. B67-B75 ◽  
Author(s):  
Aldo Leal-Egaña ◽  
Gregor Lang ◽  
Carolin Mauerer ◽  
Jasmin Wickinghoff ◽  
Michael Weber ◽  
...  

2017 ◽  
Vol 27 (36) ◽  
pp. 1701427 ◽  
Author(s):  
Jana Petzold ◽  
Tamara B. Aigner ◽  
Filip Touska ◽  
Katharina Zimmermann ◽  
Thomas Scheibel ◽  
...  

2008 ◽  
Vol 47 (24) ◽  
pp. 4592-4594 ◽  
Author(s):  
Ute K. Slotta ◽  
Sebastian Rammensee ◽  
Stanislav Gorb ◽  
Thomas Scheibel

2003 ◽  
Vol 438-439 ◽  
pp. 382-385 ◽  
Author(s):  
Luru Dai ◽  
Yong Zhang ◽  
Zhong-can Ou-Yang

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