Mirus Bio receives ISO 13485:2016 certification, underscoring the quality of processes used to support GMP product portfolio. Read more

mirus bio logo

Applications | Stem Cell Transfection

Stem Cell Transfection Solutions

Mirus Bio reagents enable high efficiency transfection of stem cells and other hard to transfect cell types used for stem cell research.

  • Perform genome editing – with TransIT-X2® Dynamic Delivery System 
  • Transfect DNA effectively – with TransIT®-2020 or TransIT®-LT1 Transfection Reagents
  • Perform repeated, low toxicity mRNA transfections – using TransIT®-mRNA Transfection Kit
  • Electroporate efficiently and cost-effectively – with Ingenio® Electroporation Solution

Poster presented at the Cellular Dynamics International User Group Meeting: High Efficiency Transfection of iCell® Cardiomyocytes and Stem Cell Relevant Cell Sources (PDF)

Optimal points of transfection for various types of stem cells illustration

Ideal Entry Points for Transfection in Stem Cell Workflow. Somatic cells such as adult fibroblast cells can be transfected or transduced via several methods (e.g. recombinant virus, plasmid, protein, mRNA, small molecule and miRNA) with a combination of transcription factors including KLF4, SOX2, c-Myc, Nanog, Oct-3/4 and LIN-28 to reprogram the cells to a pluripotent state. iPS cells can then be differentiated to a myriad of cell types through growth factor addition and/or transfection of selection markers driven by cell type specific promoters. Stem cell derived cell types such as cardiomyocytes, adipocytes, neural cells, pancreatic-β cells, and hematopoietic progenitor cells can provide researchers with relevant models for their experiments.

 

Genome Editing in Human iPSCs

CRISPR/Cas9 RNP Delivery iPSC Data

Efficient Genome Editing with Cas9 + Guide RNA Ribonucleoprotein Complexes. TransIT-X2® Dynamic Delivery System was used to deliver Cas9 protein/guide RNA ribonucleoprotein (RNP) complexes in human induced pluripotent stem cells (iPSCs). A T7E1 mismatch assay was used to measure cleavage efficiency at 48 hours post-transfection.

 

Primary and Stem Cells Transfected Using Mirus Bio Reagents

Cell TypeNucleic AcidMirus Reagent(s) UsedApplicationReference
Human embryonic stem cell derived neural progenitorsPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Human foreskin fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
Human induced pluripotent stem (iPS) cellsPlasmid DNATransIT®-2020Stem cell transfectionData
 Plasmid DNAIngenio®Stem cell transfectionData
 RNPTransIT-X2®Genome EditingData
Human mesenchymal stem cellsPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Human skin fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
iCell® CardiomyocytesPlasmid DNATransIT®-LT1Stem cell transfectionData
Mouse embryonic fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
Mouse embryonic stem cell derived cardiomyocytesPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Normal human dermal fibroblasts (NHDF)Plasmid DNATransIT®-2020DNA transfectionScientist feedback
SRF -/- mouse embryonic stem cellsPlasmid DNATransIT®-LT1Stem cell transfectionCitation
BJ human neonatal foreskin fibroblastsmRNATransIT®-mRNAStem cell reprogrammingCitation
C3H/10T1/2mRNATransIT®-mRNAStem cell reprogrammingCitation
CCD-1109Sk human normal adult skin fibroblastsmRNATransIT®-mRNAStem cell reprogrammingCitation
MRC-5 human lung fibroblastsmRNATransIT®-mRNAStem cell reprogrammingCitation
Primary human neonatal epidermal keratinocytesmRNATransIT®-mRNAStem cell reprogrammingCitation
Primary human lung fibroblastsmRNATransIT®-mRNAStem cell reprogrammingCitation
Human mesenchymal stem cellssiRNATransIT-TKO®Stem cell differentiationCitation
iCell® CardiomyocytessiRNATransIT-TKO®Knockdown of gene expressionData
Human adipose derived adult stem cells (hADAS)siRNATransIT-siQUEST®Knockdown of gene expressionCitation

DNA Transfection Into Stem Cells and Their Derivatives

TransIT®-2020 and TransIT®-LT1 Transfection Reagents for DNA Delivery

  • High Efficiency Delivery – Achieve high expression in many cell types, including hard to transfect cell types such as stem cells and primary cells
  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases
  • Suitable for Many Applications – Perform gene expression studies, promoter analysis and stable cell line generation

Download Protocol: Reverse Transfection of Human Induced Pluripotent Stem (iPS) Cells with TransIT®-LT1 Transfection Reagent (PDF)

 

TransIT-LT1 Transfection Reagent iPS Cells-Data

Exceptional Transfection Efficiency in Human Induced Pluripotent Stem Cells (iPSCs) via Reverse Transfection with TransIT®-LT1. The TransIT®-LT1 Transfection Reagent was used to reverse transfect 1.3 x 106 iPS cells with a ZsGreen expressing plasmid (Clontech). Reverse transfections were performed in 6-well plates using 12 µl of TransIT®-LT1 Transfection Reagent to deliver 4 µg of DNA (3:1, reagent: DNA). Cells were visualized 48 hours post-transfection and imaged under a 10X objective with an Olympus IX71® Inverted Microscope. Images are (A) phase contrast and (B) green fluorescence. Cells were assayed 48 hours post-transfection on an Accuri® Cytometer. The histogram (C) shows untransfected cells (black line) compared to cells transfected with plasmid using TransIT®-LT1 (green line). See more information on stem cell applications.

TransIT-LT1 Transfection Reagent mESC transfection Data

High Efficiency of Gene Expression in Mouse Embryonic Stem Cells (mESCs) using TransIT®-LT1. Mouse Embryonic Stem Cells (mESCs) were seeded at 250,000 cells per well of a 6-well plate and transfected two hours after plating with 6 μl of TransIT®-LT1 Transfection Reagent and 2.5 μg of a GFP expressing plasmid. Efficiency of transfection was visualized at approximately 60%. Images were taken using a Leica DMI 6000B inverted microscope 48 hours post-transfection. Images are (A) phase contrast, (B) green fluorescence, and (C) merged field.

Transfection of iCell CardiomyocytesData

Plasmid DNA Delivery to iCell® Cardiomyocytes Using TransIT®-LT1 Transfection Reagent. (A) High efficiency transfection of iCell® Cardiomyocytes with a GFP encoding plasmid. iCell® Cardiomyocytes were plated at 20,000 cells/well in a 96 well tissue culture plate coated with 0.1% gelatin. After allowing the cells to recover from thaw, cells were transfected with 100 ng/well of pMAXGFP (Lonza) using TransIT®-LT1 Transfection Reagent with a 2:1 reagent-to-DNA ratio according to the manufacturer’s instructions. Fluorescent images were taken 3 days post transfection using a Olympus IX71® inverted microscope. (B) cAmp induction measured via a luciferase reporter plasmid. iCell® Cardiomyocytes were plated for 5 days and subsequently replated using 40,000 or 80,000 cells/well in a 96 well plate pre-coated with gelatin. Three days post-replating cells were transfected using TransIT®-LT1 and a CRE-luciferase reporter plasmid. After 18 hours the cAMP pathway was induced using 10 µM isoproterenol for 6 hours. Luciferase activity was measured using the Promega Dual Glo® Luciferase Assay. Data is normalized to the control reporter.

Data courtesy of

 

 

Primary and Stem Cells Transfected using Mirus Bio Reagents

Cell TypeNucleic AcidMirus Reagent(s) UsedApplicationReference
Human embryonic stem cell derived neural progenitorsPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Human foreskin fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
Human induced pluripotent stem (iPS) cellsPlasmid DNATransIT®-2020Stem cell transfectionData
 Plasmid DNAIngenio®Stem cell transfectionData
Human mesenchymal stem cellsPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Human skin fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
iCell® CardiomyocytesPlasmid DNATransIT®-LT1Stem cell transfectionData
Mouse embryonic fibroblastsPlasmid DNATransIT®-2020DNA transfectionScientist feedback
Mouse embryonic stem cell derived cardiomyocytesPlasmid DNATransIT®-2020Stem cell transfectionScientist feedback
Normal human dermal fibroblasts (NHDF)Plasmid DNATransIT®-2020DNA transfectionScientist feedback
SRF -/- mouse embryonic stem cellsPlasmid DNATransIT®-LT1Stem cell transfectionCitation

RNA Transfection for Stem Cell Applications

Transfect mRNA using TransIT®-mRNA Transfection Kit for Stem Cell Reprogramming

  • Low Cellular Toxicity – Perform repeated transfections easily while maintaining cell density
  • High Efficiency Delivery – Achieve RNA delivery in hard to transfect cells including stem cells
  • Serum Compatible – Perform transfections in the presence of serum which eliminates the need for a media change and maintains cellular health
  • Deliver Various Sizes of RNA – Ideal for specialized applications, such as viral production and protein expression from mRNA
 
TransIT-mRNA BJ cells flow cytometry Data
 

TransIT®-mRNA Transfection Kit yields Higher Efficiency and Cell Viability following 14 Consecutive Transfections Compared to Lipofectamine® RNAiMAX and Stemfect™ RNA Transfection Kit. Experimental details on repeated transfections are described above. Transfection efficiency was measured by flow cytometry on a Guava® easyCyte™ 5HT following 14 consecutive daily transfections (blue bars). Cell viability was determined using cell counts measured during flow cytometry (black line grey bars). Error bars represent the standard deviation of triplicate wells.

Stem Cell Electroporation Using Ingenio® Kits

Electroporate Stem Cells Efficiently and Cost-effectively with Ingenio® Kits and Solution

  • High Efficiency – Electroporate hard to transfect cells including stem cells and their derivatives
  • Compatibility – Use with most conventional electroporation devices including Lonza-Amaxa® Nucleofector® II, Bio-Rad® or Harvard BTX®
  • Cost Savings – Reduce research budget while maximizing results

 

Protocol: Electroporation of Human Induced Pluripotent Stem (iPS) Cells with Ingenio® Electroporation Solution (PDF)

 
 

 

Plasmid DNA Electroporation of Human Induced Pluripotent Stem (iPS) Cells with Ingenio®. The Ingenio® Electroporation Kit was used to transfect 2 x 106 iPS cells on the Amaxa® Nucleofector® II Device. Cells were electroporated with 8 µg ZsGreen expressing plasmid (Clontech) in 100 µl and plated in 6-well plates at 0.33 x 106 cells/well. Cells were visualized 24 hours post-transfection and imaged under 4X objective with an Olympus IX71® Inverted Microscope. Images are (A) phase contrast, and (B) green fluorescence. Cells were also assayed 24 hours post-transfection on an Accuri® Cytometer. The histogram (C) shows unelectroporated cells (black line) compared to cells electroporated with plasmid using the Ingenio® Electroporation Kit (green line).

Data courtesy of

 

 
 

Ingenio® Kits provide a Cost-Effective and Stable Alternative to Nucleofector® Kitsa

ProductShelf LifeCost/ElectroporationbSavings
Amaxa® Nucleofector® Kit V (VCA-1003)3 months$14.64
Ingenio® Electroporation Kit (MIR 50112)1 year$8.6841%

a Compatible with Amaxa® Nucleofector® II device
b Based on 2014 U.S. List prices from company websites and protocol recommendations (25 RXN kit).

Products for Stem Cell Transfections

TransIT-X2 Transfection Reagent Sample

TransIT-X2® Dynamic Delivery System

An advanced system for delivery of CRISPR/Cas9 components in human IPSCs

  • Broad Spectrum Delivery – High efficiency delivery in a wide range of cell types
  • Genome Editing – Deliver Cas9 protein/guide RNA ribonucleoprotein (RNP) complexes
  • Technology – Novel, non-liposomal, polymeric delivery

TransIT-2020 Transfection Reagent Sample

 

TransIT®-2020 Transfection Reagent

A high performance, animal-free, broad spectrum DNA transfection reagent

  • Broad Spectrum DNA Delivery – Achieve high expression in many cell types, including hard to transfect cell lines
  • Gentle to Your Cells – Balances high efficiency nucleic acid delivery and low cellular toxicity
  • Animal Origin Free – High performance with maximum compatibility

TransIT-LT1 Transfection Reagent Sample

 

TransIT®-LT1 Transfection Reagent

A low toxicity, broad spectrum DNA transfection reagent

  • Broad Spectrum DNA Delivery – Achieve high expression in many cell types, including hard to transfect cell lines
  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases
  • High Efficiency Delivery – Achieve expression in a large population of cells

TransIT-mRNA Transfection Reagent Sample

TransIT®-mRNA Transfection Kit

A high efficiency, low toxicity transfection reagent for large RNA delivery

  • Low Cellular Toxicity – Maintain cell density and reduce experimental biases
  • High Efficiency Delivery – Achieve RNA delivery in a large population of cells
  • Serum Compatible – Perform transfections in the presence of serum which eliminates the need for a media change

Ingenio Electroporation Solution Sample

Ingenio® Electroporation Solution

High efficiency electroporation using any instrument

  • High Efficiency Electroporation – Deliver to hard to transfect cell lines and primary cells
  • Compatible with Most Conventional Electroporation Devices – Use your existing system including Lonza-Amaxa®, Bio-Rad® or Harvard BTX®
  • Save Money Without Sacrificing Performance – Available as a stand-alone solution or as part of a complete kit with cuvettes and cell droppers
 

Technical Resources

Don’t See Your Cell Type? Consult Reagent Agent® Transfection Database
Citation Database: Check if our reagents have been used by other researchers to transfect your cell type
Technical Support: Speak directly with a transfection expert

"I recently tested TransIT®-2020 and TransIT®-LT1, and both reagents worked well in terms of their efficiency at transfecting human-derived iPS cells with CRISPR constructs and a fluorescent protein reporter. Through visual inspection, transfection efficiencies with TransIT®-2020 and TransIT®-LT1 were clearly higher than with Lipofectamine® 3000."
Fedir Kiskin
University of Cambridge