Cell Culture

Pluripotent Stem Cell Culture

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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.

Human iPSC Workflow Solutions

 

Figure 1. Human iPSC Workflow Solutions. Donor fibroblasts or PBMCs can be reprogrammed into induced pluripotent stem cells (iPSCs) with the overexpression of the 4 Yamanaka transcription factors: Oct-4, Sox-2, Klf-4 and c-Myc. These human stem cells can then be expanded and differentiated into all three germ layers of the human body (Ectoderm, Endoderm and Mesoderm) following defined differentiation protocols.

 

 Human iPS Cell Lines

The European Bank of Induced Pluripotent Stem Cells (EBiSC) is a collection of high quality human iPS cells available for researchers for use in disease modelling and other forms of stem cell research. The initial collection has been generated from a wide range of donors representing specific disease backgrounds and healthy controls. EBiSC depositors have established many routine procedures for collecting, expanding and characterizing human iPS cell lines. The stem cell bank includes iPSC cell lines derived from neurodegenerative diseases (Alzheimer’s Disease, Parkinson’s Disease, Dementia, Motor Neuron Disease (ALS) - and Huntington’s Disease), eye and heart diseases, and lines from healthy control donors for age and sex matching.

Browse the Stem Cell Bank

 

 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

Application Note: Single transfection of a synthetic polycistronic self-replicative RNA yields high numbers of transgene-free human iPSCs

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.      

Simplicon is a Next Generation Integration-Free Reprogramming Technology

Figure 2. Simplicon is a Next Generation Integration-Free Reprogramming Technology. The evolution of reprogramming technologies has culminated in the development of synthetic RNA-mediated reprogramming (extreme right), representing the safest and most efficient method for iPS cell generation. Illustration adapted from Juan A. Bernal, J. of Cardiovasc. Trans. Res. (2013) 6:956–968, July 2013.

 

Part Number Item Description
SCR550 Simplicon™ RNA Reprogramming Kit (OKSG)
SCR703 Simplicon™ OKSG-cMyc RNA (OKSGM)
SCR712 TagRFP Simplicon RNA Kit
SCR713 TagGFP2 Simplicon RNA
SCR714 Human OKSG-cMyc TagRFP Simplicon RNA

 

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

Lentiviral STEMCCA Reprogrammed iPSC colonies

Figure 3. Lentiviral STEMCCA Reprogrammed iPSC colonies. Human iPS cells generated using Human STEMCCA Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit (SCR544) (A). Human iPS cells express human pluripotent markers including alkaline phosphatase (B), SSEA-4 (C), Oct-4 (D), SSEA-3 (E), TRA-1-60 (F) and TRA-1-81 (G).

 

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.

Application Note: A Serum-Free and Feeder-Free Defined Medium That Enables Weekend-Free Culture of Undifferentiated Human ES/iPS Cells

 

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.

Human pluripotent stem cells maintain proper marker expression when cultured in PluriSTEM™ Human ES/iPS Cell Medium

Figure 4. Human pluripotent stem cells maintain proper marker expression when cultured in PluriSTEM™ Human ES/iPS Cell Medium. H9 human ES cells grown in PluriSTEM™ medium for 22 passages with feeding 3 days per week express high levels of Alkaline Phosphatase (B, Pink), Oct-4 (C, Green), Nanog (D, Red), Sox-2 (E, Green) and TRA-1-60 (F, Red). Cells were counterstained with Dapi. Cells were also analyzed for Oct-4, TRA-1-60, SSEA-4 and SSEA-1 expression by flow cytometric analysis.

 

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). The 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

 

Cell Culture