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Enhanced neuronal differentiation in a three-dimensional collagen-hyaluronan matrix
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för materialkemi.
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för medicinsk biokemi och mikrobiologi.
Vise andre og tillknytning
2007 (engelsk)Inngår i: Journal of Neuroscience Research, ISSN 0360-4012, E-ISSN 1097-4547, Vol. 85, nr 10, s. 2138-2146Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Efficient 3D cell systems for neuronal induction are needed for future use in tissue regeneration. In this study, we have characterized the ability of neural stem/progenitor cells (NS/PC) to survive, proliferate, and differentiate in a collagen type I-hyaluronan scaffold. Embryonic, postnatal, and adult NS/PC were seeded in the present 3D scaffold and cultured in medium containing epidermal growth factor and fibroblast growth factor-2, a condition that stimulates NS/PC proliferation. Progenitor cells from the embryonic brain had the highest proliferation rate, and adult cells the lowest, indicating a difference in mitogenic responsiveness. NS/PC from postnatal stages down-regulated nestin expression more rapidly than both embryonic and adult NS/PC, indicating a faster differentiation process. After 6 days of differentiation in the 3D scaffold, NS/PC from the postnatal brain had generated up to 70% neurons, compared with 14% in 2D. NS/PC from other ages gave rise to approximately the same proportion of neurons in 3D as in 2D (9-26% depending on the source for NS/PC). In the postnatal NS/PC cultures, the majority of III-tubulin-positive cells expressed glutamate, -aminobutyric acid, and synapsin I after 11 days of differentiation, indicating differentiation to mature neurons. Here we report that postnatal NS/PC survive, proliferate, and efficiently form synapsin I-positive neurons in a biocompatible hydrogel.

sted, utgiver, år, opplag, sider
2007. Vol. 85, nr 10, s. 2138-2146
Emneord [en]
3D cultures, neural stem/progenitor cells, hydrogel, scaffold, neurogenesis
HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-11683DOI: 10.1002/jnr.21358ISI: 000248516700008PubMedID: 17520747OAI: oai:DiVA.org:uu-11683DiVA, id: diva2:39452
Tilgjengelig fra: 2007-10-17 Laget: 2007-10-17 Sist oppdatert: 2017-12-11bibliografisk kontrollert
Inngår i avhandling
1. Neural Stem and Progenitor Cells as a Tool for Tissue Regeneration
Åpne denne publikasjonen i ny fane eller vindu >>Neural Stem and Progenitor Cells as a Tool for Tissue Regeneration
2009 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Neural stem and progenitor cells (NSPC) can differentiate to neurons and glial cells. NSPC are easily propagated in vitro and are therefore an attractive tool for tissue regeneration. Traumatic brain injury (TBI) is a common cause for death and disabilities. A fundamental problem following TBI is tissue loss. Animal studies aiming at cell replacement have encountered difficulties in achieving sufficient graft survival and differentiation. To improve outcome of grafted cells after experimental TBI (controlled cortical impact, CCI) in mice, we compared two transplantation settings. NSPC were transplanted either directly upon CCI to the injured parenchyma, or one week after injury to the contralateral ventricle. Enhanced survival of transplanted cells and differentiation were seen when cells were deposited in the ventricle. To further enhance cell survival, efforts were made to reduce the inflammatory response to TBI by administration of ibuprofen to mice that had been subjected to CCI. Inflammation was reduced, as monitored by a decrease in inflammatory markers. Cell survival as well as differentiation to early neuroblasts seemed to be improved.

To device a 3D system for future transplantation studies, NSPC from different ages were cultured in a hydrogel consisting of hyaluronan and collagen. Cells survived and proliferated in this culturing condition and the greatest neuronal differentiating ability was seen in cells from the newborn mouse brain.

NSPC were also used in a model of peripheral nervous system injury, and xeno-transplanted to rats where the dorsal root ganglion had been removed. Cells survived and differentiated to neurons and glia, furthermore demonstrating their usefulness as a tool for tissue regeneration.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis, 2009. s. 69
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 500
Emneord
traumatic brain injury, neural stem cells, transplantation, CNS, PNS, progenitor cells, inflammation, CCI
HSV kategori
Forskningsprogram
neurokirurgi; medicinsk biokemi
Identifikatorer
urn:nbn:se:uu:diva-110095 (URN)978-91-554-7658-8 (ISBN)
Disputas
2009-12-17, B42, Husargatan 3, BMC, 09:15 (svensk)
Opponent
Veileder
Tilgjengelig fra: 2009-11-26 Laget: 2009-11-03 Sist oppdatert: 2009-11-26bibliografisk kontrollert
2. Hyaluronan Derivatives and Injectable Gels for Tissue Engineering
Åpne denne publikasjonen i ny fane eller vindu >>Hyaluronan Derivatives and Injectable Gels for Tissue Engineering
2008 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

The present work describes the preparation of hyaluronan derivatives and hydrogels with potential use in tissue engineering applications. A potentially injectable hydrogel consisting of hyaluronan and collagen was successfully used to grow neurons in vitro by encapsulation of neural stem and progenitor cells. Attempts were further made to establish a suitable modification strategy which could be used for the preparation of in vivo cross-linkable hyaluronan derivatives. The synthesis of a model substance consisting of a D-glucuronate derivative which could simplify the development of such a modification technique is described, although a new method to prepare hyaluronan derivatives was found without its use. The modification strategy involves the use of a triazine-reagent which enables the covalent attachment of hydrophilic and hydrophobic amines to hyaluronan carboxyl groups in a controlled fashion under mild conditions. Using triazine-activated amidation we synthesized an aldehyde-derivative of hyaluronan which was used to prepare gels by cross-linking with hydrazide-modified polyvinyl-alcohol. Gels were formed in less than 1 minute by mixing equal volumes of the polymer derivatives and they were subsequently used as a carrier for bone morphogenetic protein-2. An in vitro release study showed that approximately 88% of the growth factor is retained in the gel over a 4 week period. The ability to form new bone in vivo was further evaluated in an ectopic rat model by the injection of gels containing 30 µg BMP-2. Radiographic and histological examination 4 and 10 weeks after injection showed the formation of new bone without any signs of inflammation or foreign body response. Hydroxyapatite particles were further added to improve the mechanical properties of the gel, and a comparative study was conducted. This time the induced tissue consisted not only of bone, but also of interconnected cartilage and tendon, as confirmed by histology and immunohistochemistry.

sted, utgiver, år, opplag, sider
Uppsala: Universitetsbiblioteket, 2008. s. 51
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 573
Emneord
Hyaluronan, Hydrogel, Tissue Engineering, Scaffold, Minimally invasive strategy, Bone morphogenetic protein, Hydroxyapatite, Bone, Cartilage, Tendon
HSV kategori
Identifikatorer
urn:nbn:se:uu:diva-9357 (URN)978-91-554-7335-8 (ISBN)
Disputas
2008-12-05, Polhemsalen, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2008-11-14 Laget: 2008-11-14 Sist oppdatert: 2010-03-04bibliografisk kontrollert

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