High Efficiencies of Gene Transfer with Immobilized Recombinant Retrovirus: Kinetics and Optimization

Bajaj, Bharat; Lei, Pedro; Andreadis, Stelios T.

doi:10.1021/bp010039n

Cited by 57 publications

(48 citation statements)

References 44 publications

(60 reference statements)

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“…Human neonatal foreskin fibroblasts and mice dermal fibroblasts were isolated from tissues as reported elsewhere (Bajaj et al, 2001). Mice aortic smooth muscle cells (ASMCs) were isolated from mice aorta sections as described previously (Swartz et al, 2005).…”

Section: Cell Isolation and Culturementioning

confidence: 99%

Cadherin-11 is a novel regulator of extracellular matrix synthesis and tissue mechanics

Row

Liu

Alimperti

et al. 2016

Journal of Cell Science

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We discovered that Cadherin-11 (CDH11) regulates collagen and elastin synthesis, both affecting the mechanical properties and contractile function of animal tissues. Using a Cdh11-null mouse model, we observed a significant reduction in the mechanical properties [Youngs' modulus and ultimate tensile strength (UTS)] of Cdh11 −/− as compared to wild-type (WT) mouse tissues, such as the aorta, bladder and skin. The deterioration of mechanical properties (Youngs' modulus and UTS) was accompanied by reduced collagen and elastin content in Cdh11 −/− mouse tissues as well as in cells in culture. Similarly, knocking down CDH11 abolished collagen and elastin synthesis in human cells, and consequently reduced their ability to generate force. Conversely, engagement of CDH11 through homophilic interactions, led to swift activation of the TGF-β and ROCK pathways as evidenced by phosphorylation of downstream effectors. Subsequently, activation of the key transcription factors, MRTF-A (also known as MKL1) and MYOCD led to significant upregulation of collagen and elastin genes. Taken together, our results demonstrate a novel role of adherens junctions in regulating extracellular matrix (ECM) synthesis with implications for many important biological processes, including maintenance of tissue integrity, wound healing and tissue regeneration.

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Section: Cell Isolation and Culturementioning

confidence: 99%

Cadherin-11 is a novel regulator of extracellular matrix synthesis and tissue mechanics

Row

Liu

Alimperti

et al. 2016

Journal of Cell Science

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“…The immobilization of vectors to biomaterials, which has been termed substrate-mediated delivery, solid phase delivery, or reverse transfection, mimics the natural process of virus binding to extracellular matrix proteins [97,98]. Immobilization to the adhesive matrix colocalizes the vector and adhered cells [99,100], which can overcome mass transport limitations.…”

Section: Vector Immobilizationmentioning

confidence: 99%

Matrices and scaffolds for DNA delivery in tissue engineering

Laporte¹,

Shea²

2007

Advanced Drug Delivery Reviews

240

208

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Regenerative medicine aims to create functional tissue replacements, typically through creating a controlled environment that promotes and directs the differentiation of stem or progenitor cells, either endogenous or transplanted. Scaffolds serve a central role in many strategies by providing the means to control the local environment. Gene delivery from the scaffold represents a versatile approach to manipulating the local environment for directing cell function. Research at the interface of biomaterials, gene therapy, and drug delivery has identified several design parameters for the vector and the biomaterial scaffold that must be satisfied. Progress has been made towards achieving gene delivery within a tissue engineering scaffold, though the design principles for the materials and vectors that produce efficient delivery require further development. Nevertheless, these advances in obtaining transgene expression with the scaffold have created opportunities to develop greater control of either delivery or expression and to identify the best practices for promoting tissue formation. Strategies to achieve controlled localized expression within the tissue engineering scaffold will have broad application to the regeneration of many tissues, with great promise for clinical therapies.

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“…While immobilizing DNA to a substrate may seem counterintuitive as cellular internalization is necessary for expression, viral vectors have been shown to associate with the extracellular matrix to facilitate cellular binding and internalization [34,35]. The properties of the substrate are critical determinants of the interaction strength between the complexes and the surface, which subsequently affects the extent of transfection.…”

Section: Transfection On Bulk Sams On Goldmentioning

confidence: 99%

Substrate-mediated delivery from self-assembled monolayers: Effect of surface ionization, hydrophilicity, and patterning

2005

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Gene transfer has many potential applications in basic and applied sciences. In vitro, DNA delivery can be enhanced by increasing the concentration of DNA in the cellular microenvironment through immobilization of DNA to a substrate that supports cell adhesion. Substrate-mediated delivery describes the immobilization of DNA, complexed with cationic lipids or polymers, to a biomaterial or substrate. As surface properties are critical to the efficiency of the surface delivery approach, selfassembled monolayers (SAMs) of alkanethiols on gold were used to correlate surface chemistry of the substrate to binding, release, and transfection of non-specifically immobilized complexes. Surface hydrophobicity and ionization were found to mediate both DNA complex immobilization and transfection, but had no effect on complex release. Additionally, SAMs were used in conjunction with soft lithographic techniques to imprint substrates with specific patterns, resulting in patterned DNA complex deposition and transfection, with transfection efficiencies in the patterns nearing 40%. Controlling the interactions between complexes and substrates, with the potential for patterned delivery, can be used to locally enhance or regulate gene transfer, with applications to tissue engineering scaffolds and transfected cell arrays.

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High Efficiencies of Gene Transfer with Immobilized Recombinant Retrovirus: Kinetics and Optimization

Cited by 57 publications

References 44 publications

Cadherin-11 is a novel regulator of extracellular matrix synthesis and tissue mechanics

Cadherin-11 is a novel regulator of extracellular matrix synthesis and tissue mechanics

Matrices and scaffolds for DNA delivery in tissue engineering

Substrate-mediated delivery from self-assembled monolayers: Effect of surface ionization, hydrophilicity, and patterning

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