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Electrospun polyurethane scaffolds for proliferation and neuronal differentiation of human embryonic stem cells.

Artikel i vetenskaplig tidskrift
Författare Björn Carlberg
Mathilda Zetterström Axell
Ulf Nannmark
Johan Liu
Hans-Georg Kuhn
Publicerad i Biomedical materials (Bristol, England)
Volym 4
Nummer/häfte 4
Sidor 45004
ISSN 1748-605X
Publiceringsår 2009
Publicerad vid Institutionen för neurovetenskap och fysiologi, sektionen för klinisk neurovetenskap och rehabilitering
Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi
Sidor 45004
Språk en
Länkar dx.doi.org/10.1088/1748-6041/4/4/04...
Ämneskategorier Cell- och molekylärbiologi

Sammanfattning

Adult central nervous system (CNS) tissue has a limited capacity to recover after trauma or disease. Hence, tissue engineering scaffolds intended for CNS repair and rehabilitation have been subject to intense research effort. Electrospun porous scaffolds, mimicking the natural three-dimensional environment of the in vivo extracellular matrix (ECM) and providing physical support, have been identified as promising candidates for CNS tissue engineering. The present study demonstrates in vitro culturing and neuronal differentiation of human embryonic stem cells (hESCs) on electrospun fibrous polyurethane scaffolds. Electrospun scaffolds composed of biocompatible polyurethane resin (Desmopan 9370A, Bayer MaterialScience AG) were prepared with a vertical electrospinning setup. Resulting scaffolds, with a thickness of approximately 150 microm, exhibited high porosity (84%) and a bimodal pore size distribution with peaks at 5-6 and 1 microm. The mean fiber diameter was measured to approximately 360 nm with a standard deviation of 80 nm. The undifferentiated hESC line SA002 (Cellartis AB, Göteborg, Sweden) was seeded and cultured on the produced scaffolds and allowed propagation and then differentiation for up to 47 days. Cultivation of hESC on electrospun fibrous scaffolds proved successful and neuronal differentiation was observed via standard immunocytochemistry. The results indicate that predominantly dopaminergic tyrosine hydroxylase (TH) positive neurons are derived in co-culture with fibrous scaffolds, in comparison to reference cultures under the same differentiation conditions displaying large proportions of GFAP positive cell types. Scanning electron micrographs confirm neurite outgrowth and connection to adjacent cells, as well as cell attachment to individual fibers of the fibrous scaffold. Consequently, electrospun polyurethane scaffolds have been proven feasible as a substrate for hESC propagation and neuronal differentiation. The physical interaction between cells and the fibrous scaffold indicates that these scaffolds provide a three-dimensional physical structure; a potential candidate for neural tissue engineering repair and rehabilitation.

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