Cell Culture

Pluripotent Stem Cell Culture

Embryonic and Induced Pluripotent Stem Cells

Pluripotent stem cells, including embryonic stem cells (ES cells) and induced pluripotent stem cells (iPSCs), have the capacity to give rise to differentiated progeny representative of all three germ layers (ectoderm, endoderm, and mesoderm). The ability to expand pluripotent cells in vitro and subject them to direct differentiation to produce specific cell types is crucial to the development of cell-based therapies to replace or restore tissue that has been damaged by disease or injury.

The discovery that somatic cells could revert back to pluripotent like cells (induced pluripotent stem cells) in 2006 by Shinya Yamanaka created an entirely new area of stem cell biology. IPS cells removed the ethical concerns associated with embryonic stem cells and allowed scientists to model human diseases that were previously impossible to model such as Alzheimer’s, Parkinson’s and Autism.

MilliporeSigma offers a broad range of tools to support the iPS workflow including: iPS cell reprogramming kits; iPS cell culture media and reagents; iPS cell characterization tools; and iPS cell differentiation media. Our range of ES cell-qualified products provides researchers with convenient and cost effective solutions for the reliable culture of pluripotent ES/iPS cells.

Current iPSC Workflow

1. Takahashi et al. (2007)
2. Yu et al. (2007)

Illustration adapted from Alexandra Rolletschek and Anna M. Wobus Biol. Chem., Vol. 390, pp. 845–849, September 2009.


Pluripotent stem cells have the ability to differentiate into all three germ layers (mesoderm, ectoderm and endoderm)

 

iPSC Reprogramming Kits

Virus-Free iPS Reprogramming Kits

The Simplicon™ RNA Reprogramming Technology utilizes a single polycistronic self-replicating RNA strand that is sufficient for generating high numbers of human iPSCs using a single transfection step. The single RNA contains the four reprogramming factors, OCT-4, KLF-4, SOX-2 and GLIS1, and allows for efficient reprogramming without viral intermediates or host genome integration. 

With Simplicon™ RNA Reprogramming Technology, you can:

  • Create integration-free iPS cells using synthetic RNA
  • Reprogram efficiently and easily using a single transfection step
  • Achieve controlled elimination of reprogramming factors

Watch Simplicon Video Here

See the Data

Technology Featured in Cell Stem Cell: Yoshioka, Naohisa, et al. Efficient generation of human iPSCs by a synthetic self-replicative RNA. Cell Stem Cell 13.2 (2013): 246-254.      

 

Part Number Item Description
SCR550 Simplicon™ RNA Reprogramming Kit (OKSG)
SCR549 Simplicon™ Reprogramming RNA (OKSG)
SCR703 Simplicon™ OKSG-cMyc RNA (OKSGM)
GF156 B18R, Human Recombinant Carrier-Free
SCM094 Human iPS Reprogramming Boost Supplement II
TR-1013 RiboJuice™ mRNA Transfection Kit

 

Lentiviral iPS Reprogramming Kits

The STEMCCA™ lentivirus reprogramming kits make it easier than ever to obtain and work with induced pluripotent stem (iPS) cells even with the most difficult to reprogram cells such as PBMCs. Unlike traditional iPS cell generation which requires simultaneous co-infection by four separate expression vectors, the STEMCCA kits use a single polycistronic lentiviral vector to improve efficiency and reduce the number of viral integrations.

See the Data

Learn how our STEMCCA lentiviral reprogramming system can be used to efficiently generate iPS cells from 3-4ml of peripheral blood

Ordering Info
 

Part Number Item Description
SCR545 Human STEMCCA™ Cre-Excisable Constitutive
SCR548 Human STEMCCA Cre-Excisable Constitutive Polycistronic (OKS/L-Myc)
SCR530 Mouse STEMCCA™ Constitutive Polycistronic
SCR531 Mouse STEMCCA™ Cre-Excisable Constitutive
SCR508 TAT-Cre Recombinase
TR-1003 Polybrene Infection / Transfection Reagent

 

Reprogramming Boost Supplement Kits

Traditionally, iPS cell generation has been plagued by low reprogramming efficiencies depending on the method of iPS cell generation and starting cell type used. Millipore has developed various reprogramming boost kits which contain a collection of validated small molecule inducers and inhibitors to increase the overall reprogram efficiencies achieved with any reprogramming method used.

Reprogramming Boost Kit Advantages:

  • Decreases time required to establish fully reprogrammed colonies by 50%
  • Enhances generation of fully reprogrammed colonies
  • Improves colony morphology — colonies possess distinctive flat 2D morphology and can be easily passaged
  • More than doubles reprogramming efficiency
Part Number Item Description Inhibitors Included
SCM087 Mouse iPS Reprogramming Boost Supplement TGF-ß RI Kinase Inhibitor II
GSK3ß Inhibitor
Methylation Modulator
SCM088 Human iPS Reprogramming Boost Supplement I TGF-ß RI Kinase Inhibitor VI
HDAC Inhibitor-A
SCM094 Human iPS Reprogramming Boost Supplement II TGF-β RI Kinase Inhibitor IV
Sodium Butyrate
PS48

 

 Pluripotent Stem Cell Culture Media

PluriSTEM™ Human ES/iPS Medium
Feed Less. Discover More

MilliporeSigma’s PluriSTEM™ Human ES/iPS Medium is a specially formulated defined media that maintains human pluripotent stem cells in feeder-free and serum-free conditions with less frequent feeding and cell culture time. The proprietary formulation utilizes Activin-A, TGFβ1, and b-FGF to promote stem cell self-renewal and potent small molecule combinations to inhibit unwanted spontaneous differentiation.

See the Data

Features and Benefits

  • Feeder-Free, Serum-Free, Defined Cell Culture System for Human ES/iPS Cells.
  • Less Frequent Cell Feeding (Every other day, weekend free) Reducing Cost and Overall Cell Culture Time.
  • Small Molecule based media with Low Total Protein and Growth Factor Concentrations.
  • High Viability and Proliferation in Single Cell Passaging.

Ordering Info
 

Part Number Item Description
SCM130 PluriSTEM™ Human ES/iPS Medium
SCM132 PluriSTEM-XF™ Human ES/iPS Medium
SCM133 PluriSTEM™ Dispase-II Solution
SCM134 PluriSTEM™ Freeze Medium
SCM135 PluriSTEM-XF™ Freeze Medium
CC130 PluriSTEM-XF™ Recombinant Vitronectin

 

 Pluripotent Stem Cell Markers

Pluripotent Stem Cell Antibodies

Browse our extensive selection of human pluripotent stem cell specific antibodies including OCT-4, SOX-2, Lin-28, TRA-1-60, TRA-1-81, SSEA-1, SSEA-3, SSEA-4, Human Nuclei and many others.

Popular Targets

Oct-4   Nanog   Sox-2   TRA-1-60   TRA-1-81   SSEA-1   SSEA-3   SSEA-4   Nestin   Human Nuclei

Browse all of our Stem Cell Antibodies Here

Pluripotent Stem Cell Characterization Kits

A convenient collection of stem cell validated primary and secondary antibodies, enzymatic stains and dyes to characterize your ES/IPS cell’s pluripotent state.

The undifferentiated state of human and mouse ES cells can be characterized by high level of expression of Alkaline Phosphatase (AP). MilliporeSigma’s Alkaline Phosphatase Detection Kit is a specific and sensitive tool for the phenotypic assessment of ES cell differentiation by the determination of AP activity.




 

 

Part Number Item Description
SCR001 ES Cell Characterization Kit
SCR002 ES Cell Marker Sample Kit
SCR077 Fluorescent Mouse ES/iPS Cell Characterization Kit
SCR078 Fluorescent Human ES/iPS Cell Characterization Kit
FCSC100107 FlowCellect™ Human iPS Cell Characterization Kit
SCR004 Alkaline Phosphatase Detection Kit
SCR066 Quantitative Alkaline Phosphatase ES Characterization Kit
SCR502 Human, iPS Selection Kit