Competitive electroporation formulation for cell therapy

Flanagan, Michael; Gimble, Jeffrey M.; Yu, Gang; Wu, Xiaoyan; Xia, Xueqing; Hu, Jiemiao; Yao, Shaomian; Li, Shulin

doi:10.1038/cgt.2011.27

Cited by 21 publications

(25 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since Flanagan and colleagues have shown that certain polymers can boost the transfection efficiency for some cells [34], we asked whether different amounts of polyethylene glycol (PEG - 0.01, 0.1 and 1%) added to 2S buffer during electroporation could augment the number of viable cells and transgene expression in mouse T cells. We observer no positive impact on cell viability whereas slight variations in GFP expression were observed; however, solution 2S achieved higher percentages of GFP positive cells in the absence of PEG (Figure S4).…”

Section: Resultsmentioning

confidence: 99%

An Efficient Low Cost Method for Gene Transfer to T Lymphocytes

et al. 2013

View full text Add to dashboard Cite

Gene transfer to T lymphocytes has historically relied on retro and lentivirus, but recently transposon-based gene transfer is rising as a simpler and straight forward approach to achieve stable transgene expression. Transfer of expression cassettes to T lymphocytes remains challenging, being based mainly on commercial kits.AimsWe herein report a convenient and affordable method based on in house made buffers, generic cuvettes and utilization of the widely available Lonza nucleofector II device to promote efficient gene transfer to T lymphocytes.ResultsThis approach renders high transgene expression levels in primary human T lymphocytes (mean 45%, 41–59%), the hard to transfect murine T cells (mean 38%, 36–42% for C57/BL6 strain) and human Jurkat T cell line. Cell viability levels after electroporation allowed further manipulations such as in vitro expansion and Chimeric Antigen Receptor (CAR) mediated gain of function for target cell lysis.ConclusionsWe describe here an efficient general protocol for electroporation based modification of T lymphocytes. By opening access to this protocol, we expect that efficient gene transfer to T lymphocytes, for transient or stable expression, may be achieved by an increased number of laboratories at lower and affordable costs.

show abstract

Section: Resultsmentioning

confidence: 99%

An Efficient Low Cost Method for Gene Transfer to T Lymphocytes

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Following our optimized nucleofection method developed previously [10], we initially chose two buffers: OptiMEM and pulsing buffer. To determine the nucleofection program that would yield the highest effectiveness, we used the following seven programs, as outlined in the Amaxa Nucleofector Optimization Protocol: A-20, T-20, T-30, X-01, X-05, L-29, and D-23.…”

Section: Resultsmentioning

confidence: 99%

Competitive DNA transfection formulation via electroporation for human adipose stem cells and mesenchymal stem cells

Flanagan

Gimble

et al. 2012

Biol Proced Online

View full text Add to dashboard Cite

BackgroundAdipose stem cells have a strong potential for use in cell-based therapy, but the current nucleofection technique, which relies on unknown buffers, prevents their use.ResultsWe developed an optimal nucleofection formulation for human adipose stem cells by using a three-step method that we had developed previously. This method was designed to determine the optimal formulation for nucleofection that was capable of meeting or surpassing the established commercial buffer (Amaxa), in particular for murine adipose stem cells. By using this same buffer, we determined that the same formulation yields optimal transfection efficiency in human mesenchymal stem cells.ConclusionsOur findings suggest that transfection efficiency in human stem cells can be boosted with proper formulation.

show abstract

“…ADSC were nucleofected with a plasmid coding for a protein involved in angiogenesis and cell proliferation: Sonic Hedgehog (Shh). In this study, the choice of nucleofection method of Shh gene was based on two arguments: (1) this technique allows the genetic material to be delivered directly into the nucleus [114,115], resulting in higher transfection efficiency than electroporation [116], (2) this technique does not alter the phenotype and potential multilineage of ADSC [117]. In this in vivo study, we checked that nucleofection technique allowed the production of sufficient amounts of transfected ADSC to be grafted to a large animal model [113].…”

Section: Stem Cell Therapy Strategies Under Assessmentmentioning

confidence: 99%

Advances in Stem Cell Therapy: Specific Applications in the Treatment of Cutaneous Radiation Syndrome

Riccobono¹,

François²

2014

J Stem Cell Res Ther

View full text Add to dashboard Cite

The emergence of stem cell therapy and cellular engineering during the last 10 years has allowed new therapeutic strategies to be considered for the treatment of many wound repair disorders in bones, muscles and skin. The cutaneous radiation syndrome that is often associated with radiation burn may also take advantage of these scientific advances. This syndrome, characterized by inflammatory waves, incomplete wound healing and poor revascularization is the dramatic consequence of local irradiation exposure (above 15Gy). Until the late 90's, the treatment schedule which exhibited poor efficacy consisted in excision followed by transient coverage of the wound bed and then by autologous skin grafts. Recently the local injection of autologous bone marrow mesenchymal stem cells, which favor wound healing and reduce pain, has achieved a major advance. However this strategy hampered by culture delays and requiring non-irradiated areas to harvest stem cells remains to be optimized. Autologous or allogeneic adipose tissue-derived stem cells, easy to collect and expand, may represent a valuable therapeutic alternative especially through pro-angiogenic and anti-inflammatory properties. Other proposed strategies including stem cell manipulation to produce trophic factors (transient gene therapy), bone marrow mesenchymal stem cells or adipose tissue derived stem cells culture media injection appear as valuable alternatives. In this review we report the latest scientific progresses in pre-clinical and clinical studies concerning stem cell therapy for cutaneous radiation syndrome.

show abstract

Competitive electroporation formulation for cell therapy

Cited by 21 publications

References 47 publications

An Efficient Low Cost Method for Gene Transfer to T Lymphocytes

An Efficient Low Cost Method for Gene Transfer to T Lymphocytes

Competitive DNA transfection formulation via electroporation for human adipose stem cells and mesenchymal stem cells

Advances in Stem Cell Therapy: Specific Applications in the Treatment of Cutaneous Radiation Syndrome

Contact Info

Product

Resources

About