A microneedle array able to inject tens of thousands of cells simultaneously

Teichert, Gregory H.; Burnett, Sandra H.; Jensen, Brian D.

doi:10.1088/0960-1317/23/9/095003

Cited by 17 publications

(26 citation statements)

References 37 publications

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“…Delivery efficiencies for these experiments ranged from 25-78%, depending upon PI concentration and application of voltage. Dif fusion alone has previously produced efficiencies as high as 98% at much higher PI concentrations [22]. PI delivery efficiency increased with increasing PI concentration.…”

Section: Resultsmentioning

confidence: 99%

“…Each 2 x 2 cm silicon chip contains approximately 4 x 106 lances on the surface, with the space between lances being 10,um. We optimized lance geometry and spacing for testing on a culture of HeLa 229 cells [22],…”

Section: Lance Array Fabrication and Injection Devicementioning

confidence: 99%

“…We have previously developed a nanoinjection lance array for injecting thousands of culture cells simultaneously [22]. We have used PI as the injection particle to measure particle uptake effi ciency and cell viability when using the nanoinjection lance array.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Injection of Propidium Iodide into HeLa Cells Using a Silicon Nanoinjection Lance Array

Lindstrom

Brewer

Ferguson

et al. 2014

Journal of Nanotechnology in Engineering and Medicine

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Delivering foreign molecules into human cells is a wide and ongoing area of research. Gene therapy, or delivering nucleic acids into cells via nonviral or viral pathways, is an especially promising area for pharmaceutics. All gene therapy methods have their respective advantages and disadvantages, including limited delivery efficiency and low viability. We present an electromechanical method for delivering foreign molecules into human cells. Nanoinjection, or delivering molecules into cells using a solid lance, has proven to be highly efficient while maintaining high viability levels. This paper describes an array of solid silicon microlances that was tested to determine efficiency and viability when nanoinjecting tens of thousands of HeLa cells simultaneously. Propidium iodide (PI), a dye that fluoresces when bound to nucleic acids and does not fluoresce when unbound, was delivered into cells using the lance array. Results show that the lance array delivers PI into up to 78% of a nanoinjected HeLa cell culture, while maintaining 78-91 % viability. With these results, we submit the nanoinjection method using a silicon lance array as another promising particle delivery method for mammalian culture cells.

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Section: Resultsmentioning

confidence: 99%

Section: Lance Array Fabrication and Injection Devicementioning

confidence: 99%

See 1 more Smart Citation

Injection of Propidium Iodide into HeLa Cells Using a Silicon Nanoinjection Lance Array

Lindstrom

Brewer

Ferguson

et al. 2014

Journal of Nanotechnology in Engineering and Medicine

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“…More specifically, first‐generation nanoinjection consisted of a micro‐electromechanical system (MEMS) that uses electrostatic attraction of DNA onto a silicon micro‐sized solid lance that is then inserted into mouse embryos before electrically releasing exogenous DNA into the host . This same technology has been modified in a second generation to consist of a microfabricated silicon wafer that has a grid of four million 10‐micron‐length lances that allow for effective delivery of genetic loads to hundreds of thousands of cells simultaneously . As an emerging method, nanoinjection shows promise in regard to in vitro CRISPR‐Cas9 chronic wound applications, given the high efficiency of transfection and high cell survival rates as well as the lack of cytotoxic effects.…”

Section: Emerging Conceptsmentioning

confidence: 99%

A review of genetic engineering biotechnologies for enhanced chronic wound healing

Sessions

Armstrong

Hope

et al. 2017

Experimental Dermatology

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Traditional methods for addressing chronic wounds focus on correcting dysfunction by controlling extracellular elements. This review highlights technologies that take a different approach -enhancing chronic wound healing by genetic modification to wound beds. Featured cutaneous transduction/transfection methods include viral modalities (ie adenoviruses, adeno-associated viruses, retroviruses and lentiviruses) and conventional non-viral modalities (ie naked DNA injections, microseeding, liposomal reagents, particle bombardment and electroporation). Also explored are emerging technologies, focusing on the exciting capabilities of wound diagnostics such as pyrosequencing as well as site-specific nuclease editing tools such as CRISPR-Cas9 used to both transiently and permanently genetically modify resident wound bed cells.Additionally, new non-viral transfection methods (ie conjugated nanoparticles, multielectrode arrays, and microfabricated needles and nanowires) are discussed that can potentially facilitate more efficient and safe transgene delivery to skin but also represent significant advances broadly to tissue regeneration research. K E Y W O R D Schronic wound healing, CRISPR-Cas9, pyrosequencing, transfection nanotechnology, viral transduction 1

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“…LAN uses a microfabricated silicon etched array of lances to physically penetrate hundreds of thousands of cells simultaneously and electrically deliver attached molecular loads (Lindstrom et al 2014; Sessions et al 2014; Teichert et al 2013) (see Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

et al. 2016

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BackgroundCRISPR-Cas9 genome editing and labeling has emerged as an important tool in biologic research, particularly in regards to potential transgenic and gene therapy applications. Delivery of CRISPR-Cas9 plasmids to target cells is typically done by non-viral methods (chemical, physical, and/or electrical), which are limited by low transfection efficiencies or with viral vectors, which are limited by safety and restricted volume size. In this work, a non-viral transfection technology, named lance array nanoinjection (LAN), utilizes a microfabricated silicon chip to physically and electrically deliver genetic material to large numbers of target cells. To demonstrate its utility, we used the CRISPR-Cas9 system to edit the genome of isogenic cells. Two variables related to the LAN process were tested which include the magnitude of current used during plasmid attraction to the silicon lance array (1.5, 4.5, or 6.0 mA) and the number of times cells were injected (one or three times).ResultsResults indicate that most successful genome editing occurred after injecting three times at a current control setting of 4.5 mA, reaching a median level of 93.77 % modification. Furthermore, we found that genome editing using LAN follows a non-linear injection-dose response, meaning samples injected three times had modification rates as high as nearly 12 times analogously treated single injected samples.ConclusionsThese findings demonstrate the LAN’s ability to deliver genetic material to cells and indicate that successful alteration of the genome is influenced by a serial injection method as well as the electrical current settings.

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A microneedle array able to inject tens of thousands of cells simultaneously

Cited by 17 publications

References 37 publications

Injection of Propidium Iodide into HeLa Cells Using a Silicon Nanoinjection Lance Array

Injection of Propidium Iodide into HeLa Cells Using a Silicon Nanoinjection Lance Array

A review of genetic engineering biotechnologies for enhanced chronic wound healing

CRISPR-Cas9 directed knock-out of a constitutively expressed gene using lance array nanoinjection

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