Efficient expansion and gene transduction of mouse neural stem/progenitor cells on recombinant fibronectin

Rappa, Germana; Kunke, David; Holter, JC; Diep, Dzung B.; Meyer, Johann; Baum, Christopher; Fodstad, Øystein; Krauß, Stefan; Lorico, Aurelio

doi:10.1016/j.neuroscience.2003.11.030

Cited by 35 publications

(32 citation statements)

References 28 publications

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“…When used to facilitate NPC retroviral transduction, it increases NPC proliferation in the neonatal mouse (Rappa et al, 2004). In contrast, in the dog we found that fibronectin did not enhance growth rate or expansion of OB-cNPCs.…”

Section: Discussioncontrasting

confidence: 63%

In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

Walton

Wolfe

2008

Journal of Neuroscience Methods

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The dog serves as a large animal model for multiple neurologic diseases that may potentially benefit from neural progenitor cell (NPC) transplantation. In the adult brain, multipotent NPCs reside in the subventricular zone and its rostral and caudal extensions into the olfactory bulb and hippocampus. The olfactory bulb represents a surgically accessible site for obtaining cells for autologous NPC transplantation. To model conditions that would occur for ex vivo gene therapy in the postnatal brain, NPCs were isolated from the canine olfactory bulb, expanded ex vivo under different culture conditions, and compared quantitatively for growth and immunophenotype. Under standard growth conditions, canine olfactory bulb-derived NPCs (OB-cNPCs) could be expanded nearly 500-fold in the time evaluated. Canine OB-cNPCs grown on poly-D-lysine (PDL) or on PDL-fibronectin had similar growth rates, whereas supplementation with leukemia inhibitory factor (LIF) resulted in significantly slower growth. However, when OB-cNPC cultures were grown on PDL-fibronectin or PDL supplemented with LIF, a greater proportion of cells with neuronal markers were generated upon differentiation.

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Section: Discussioncontrasting

confidence: 63%

In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

Walton

Wolfe

2008

Journal of Neuroscience Methods

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“…Stem cell therapies have received increased attention in regenerative medicine [189][190][191][192][193][194][195][196][197][198][199][200][201]. The use of cellular systems inside nerve conduits is intended to promote axon regeneration [202][203][204][205][206].…”

Section: Cellular Systemsmentioning

confidence: 99%

Scaffolds for Peripheral Nerve Regeneration, the Importance of In Vitro and In Vivo Studies for the Development of Cell-Based Therapies and Biomaterials: State of the Art

Pedrosa¹,

Caseiro²,

Santos³

et al. 2017

Scaffolds in Tissue Engineering - Materials, Technologies and Clinical Applications

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Human adult peripheral nerve injuries are a high incidence clinical problem that greatly affects patients' quality of life. Although peripheral nervous system has intrinsic regenerative capacity, this occurs in an incomplete or poorly functional manner. When a nerve fiber loses its continuity with consequent damage of the basal lamina tubes, axon spontaneous regeneration is disorganized and mismatched. These phenomena translate in an inadequate nerve functional recovery and consequent musculoskeletal incapacity. Nerve grafts still remain the gold standard in peripheral injuries treatment. However, this approach contains its disadvantages such as the necessity of primary surgery to harvest the autografts, loss of a functional nerve, donor site morbidity and longer surgery procedures. Therefore, biomaterials and tissue engineering can provide efficient resources and alternatives to nerve injury repair not only by the development of biocompatible structures but also, introducing neurotrophic factors and cellular systems to stimulate optimum clinical outcome. In this chapter, a comprehensive state-of-the art picture of tissue-engineered nerve grafts scaffolds, their application in nerve regeneration along with latest advances in peripheral nerve repair and future perspectives will be discussed, including our own large experience in this field of knowledge.

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“…10 Under protocol conditions, NSCs grow faster on rFN matrix than as neurospheres and do not lose their stem cell nature or multipotentiality; our protocol allows to transduce up to 90% NSCs at a multiplicity of infection (MOI) of 2 with no need for viral concentration or production of serum-free retroviral supernatants. 9 Our studies have been the first to report gene transfer by an FMEV-derived backbone in NSCs: the FMEV retroviral vectors were optimized for efficient transgene expression in hematopoietic progenitor cells, and we found them similarly well suited for NSCs and derived cell types. 9 In the present study, we have employed this retroviral-based transduction protocol to arm NSCs with the antiangiogenic molecule, endostatin or with the human prodrug-activating cytochrome P450 enzyme 2B6.…”

Section: Introductionmentioning

confidence: 74%

“…9 Our studies have been the first to report gene transfer by an FMEV-derived backbone in NSCs: the FMEV retroviral vectors were optimized for efficient transgene expression in hematopoietic progenitor cells, and we found them similarly well suited for NSCs and derived cell types. 9 In the present study, we have employed this retroviral-based transduction protocol to arm NSCs with the antiangiogenic molecule, endostatin or with the human prodrug-activating cytochrome P450 enzyme 2B6.…”

Section: Introductionmentioning

confidence: 74%

“…Primary NSCs are generally grown as floating cultures, to form heterogeneous structures comprised of undifferentiated cells and their progenies, called neurospheres. We have recently developed a highly efficient cell culture protocol 9 based on adherent culture of NSCs from neonatal forebrains on recombinant fibronectin (rFN). rFN facilitates retroviral infection by co-localization of hematopoietic cells and viral particles on the same rFN molecule.…”

Section: Introductionmentioning

confidence: 99%

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Primary neural stem/progenitor cells expressing endostatin or cytochrome P450 for gene therapy of glioblastoma

et al. 2008

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Despite recent technical improvements in surgical excision techniques and adjuvant radio-and chemotherapy, the clinical outcome of patients with grade IV astrocytoma (glioblastoma) remains very poor, with a median survival of less than 12 months. A promising approach to therapy employs gene-engineered neural stem/progenitor cells (NSCs) as a cellular therapeutic delivery system, to track glioblastoma cells and deliver anticancer molecules. However, most results on their tumor tropism have been derived by immortalized NSCs. We now report that primary murine gene-engineered NSCs displayed in vivo tropism for glioblastoma cells. Ten days after injection into the brain, many NSCs continued to express the transgene and the NSC marker, nestin. NSCs transduced with a retroviral vector co-expressing a secretable form of human endostatin and eGFP (NSC/endo-eGFP) released potentially antiangiogenetic concentrations of endostatin into the culture medium. Conditioned medium of NSC/endoeGFP cells inhibited the growth of mouse pulmonary microvascular endothelial cells (PMVECs). A good correlation between endostatin levels and PMVEC growth inhibition was observed. In NSCs co-expressing cytochrome P450 2B6 (CYP2B6) and eGFP (NSC/CYP2B6-eGFP), the forced expression of CYP2B6 resulted in intracellular activation of CPA and subsequent cell death. In the presence of NSC/CYP2B6-eGFP, we observed CPA cytotoxicity to DsRed-expressing U87Mg glioma cells. In vivo treatment of intracranial GL-261 glioblastoma with NSC/endo-eGFP caused a 65% reduction in tumor size, compared to untreated control mice or mice treated with NSC/eGFP cells. Our data suggest that primary NSCs transduced with retroviral vectors expressing endostatin and/or CYP2B6 have a potential role in glioblastoma therapy.

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Efficient expansion and gene transduction of mouse neural stem/progenitor cells on recombinant fibronectin

Cited by 35 publications

References 28 publications

In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

In vitro growth and differentiation of canine olfactory bulb-derived neural progenitor cells under variable culture conditions

Scaffolds for Peripheral Nerve Regeneration, the Importance of In Vitro and In Vivo Studies for the Development of Cell-Based Therapies and Biomaterials: State of the Art

Primary neural stem/progenitor cells expressing endostatin or cytochrome P450 for gene therapy of glioblastoma

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