Transfection into iPS Cells / ES Cells / Other Stem Cells by Electroporation

Transfection into iPS Cells / ES Cells / Other Stem Cells by Electroporation

APPLICATIONS

Transfection Results with Nepa Gene Electroporation System

Human iPS Cells

3 days after electroporation

Human iPS Cells

2 days after electroporation

7 days after electroporation

GFP is still expressed in the iPS cell colonies after cell passaging.

Mouse ES Cells

Viability: 74%

Transfection Efficiency:  88%

Mouse Neurospheres

Viability: 90%

Transfection Efficiency:  75%

Embryoid Body in Adherence from human iPS Cells

 

 

Generation of Induced Pluripotent Stem Cells (iPSCs)

Data of disease-specific iPSC Generation (disease: LQT)

Transfection of multiple episomal plasmids into B cells

iPSC colony one month after electroporation. (5X objective)

Alkaline phosphatase stain (Left: 4X objective Right: 10X objective)

Good expressions of stem cell markers TRA1-60 and OCT4.

(left green: TRA1-60, right green: OCT4, blue: DAPI, red: Phalloidin)

Courtesy of Dr. Toshio Nakanishi laboratory, Department of Pediatric Cardiology, Tokyo Women’s Medical University, Japan

 

Evaluation of CRISPR activity of human iPS cells with NEPA21 transfection

NEPA21 ips

  1. Optimization of transfection conditions using the NEPA21 super electroporator (Nepa Gene Co., Ltd.) in human iPS cell lines. Voltage and pulse width of Poring Pulse were examined. EGFP expression plasmids were transfected and the percentage of positive cells was analyzed using the LSRFortessa Cell Analyzer (BD).
  2. Schematic of transfection procedure.
  3. Cas9 expression vector pHL-EF1a-SphcCas9-iP-A (Addgene ID: 60599) and sgRNA expression vector pHL-H1-ccdB-mEF1a-RiH (Addgene ID: 60601) were electroporated into iPS cells and then the endogenous dystrophin gene cleavage activity was evaluated by the T7EI assay. The intensity of the cleaved bands (◀ red arrows) was detected with a TapeStation (Agilent) and the values are shown below the gel electrogram images.
  4. The same genomic DNA samples as in (C) were analyzed by RFLP with restriction enzyme XcmI (50-CCANNNNN^NNNTGG-30). The intensity of the uncut bands is indicated by ◀ red arrows. Because gRNA5 cleaves far from the XcmI site, no cleavage activity was detected in the RFLP assay.

Courtesy of Dr. HongMei Li and Dr. Akitsu Hotta, Center for iPS Cell Research and Application (CiRA), Kyoto University

 

Transfection Data: iPS Cells / ES Cells / Other Stem Cells

See the cell images by clicking the cell names.

V: Viability, TE: Transfection Efficiency.

Cells nameVTE Cells nameVTE
Human iPS Cells (201B7)86%70% Human iPS Cells94%80%
Human iPS Cells   Human iPS Cells  
Human iPS Cells (201B7)85%94% Human iPS Cells (201B7)  
Human iPS Cells69%80% Human iPS Cells 73%
Human iPS Cells Derived Neural Cells93%54%    
Human ES Cells (H9 p.51)55%55% Human ES Cells  
Human Mesenchymal Stem cells (Primary)78%75%  Human Mesenchymal Stem Cells70%80%
Human Neural Stem Cells97%95% Human Neural Stem Cells80%83%
Human Deciduous Teeth Stem Cells (SHED)90%92% Human Nucleated Cells Including Hematopoietic Stem Cells (Before cell isolation)73%90%
Mouse iPS Cells70%50% Mouse ES Cells80%75%
Mouse ES Cells80%68% Mouse ES Cells 74%88%
Mouse ES cells (129 strain, R1/E) 80%90% Mouse ES Cells70%100%
Mouse ES Cells80%90% Mouse iPS cell derived Neural Stem Cells 86%
Mouse Neural Stem Cells 90%80% Mouse Neural Stem cells (Primary)80%60%
Mouse Neurospheres90%75% Mouse Neurospheres  
Mouse Trophoblast Stem Cells59%47% C3H/10T1/2 Mouse Mesenchymal Stem Cells 70%85%
Mouse Mesenchymal Stem Cells99%89% Mouse Hematopoietic Stem Cells (c-Kit positive cells)66%45%
Rat ES Cells70%76% Rat ES Cells60%80%

We have a lot of data of iPS/ES/Other stem cells transfection with high efficiency and high viability. Please feel to free to contact us for the latest data.

 

PUBLICATIONS

Electroporation

Drug Delivery and Transfection

Electro Cell Fusion

Fluorescent Staining

Single-Cell/Micro-Particle Transfer

Cell Freezing

Mechanical Vibration