Transfection into iPS/ES/Other Stem Cells by Electroporation

Transfection into iPS/ES/Other Stem Cells by Electroporation

APPLICATIONS

Transfection into iPS & ES Cells

Human iPS Cells

3 days after electroporation

Human iPS Cells

2 days after electroporation and transfection into the colony

7 days after electroporation (The picture above shows the GFP expression of the iPS colony after the passage.)

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)

The pictures below show 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 by 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 into iPS/ES/Other Stem Cells

See the cell images by clicking the cell names.

V: Viability, TE: Transfection Efficiency.

Cells name V TE   Cells name V TE
Human iPS Cells (201B7) 86% 70%   Human iPS Cells 94% 80%
Human iPS Cells       Human iPS Cells    
Human iPS Cells (201B7) 85% 94%   Human iPS Cells (201B7)    
Human iPS cells 69% 80%   Human iPS Cells   73%
Human ES cells (H9 p.51) 55% 55%   Human ES Cells    
Human Mesenchymal Stem cells (Primary) 78% 75%    Human Mesenchymal Stem Cells 70% 80%
Human Neural Stem Cells 97% 95%   Human Neural Stem Cells 80% 83%
Human Deciduous Teeth Stem Cells (SHED) 90% 92%   Human Nucleated Cells Including Hematopoietic Stem Cells (Before cell isolation) 73% 90%
Mouse iPS Cells 70% 50%   Mouse ES Cells 80% 75%
Mouse ES Cells 80% 68%   Mouse ES Cells  74% 88%
Mouse ES cells (129 strain, R1/E)  80% 90%   Mouse ES Cells 70% 100%
Mouse ES Cells 80% 90%   Mouse iPS cell derived Neural Stem Cells   86%
Mouse Neural Stem Cells  90% 80%   Mouse Neural Stem cells (Primary) 80% 60%
Mouse Neurospheres 90% 75%   Mouse Neurospheres    
Mouse Trophoblast Stem Cells 59% 47%   C3H/10T1/2 Mouse Mesenchymal Stem Cells  70% 85%
Mouse Mesenchymal Stem Cells 99% 89%   Mouse Hematopoietic Stem Cells (c-Kit positive cells) 66% 45%
Rat ES Cells 70% 76%   Rat ES Cells 60% 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