* SBI is an Authorized and Fully Licensed provider of the Minicircle Reprogramming DNA Kit and the mc-iPS Cell Line.
Nonviral Human iPS Cells and Minicircle Reprogramming DNA
Nonviral and non-integrating minicircle (mc) DNA technology available as well as a Human mc-iPS Cell Line
The minicircle DNA features easy-to-transfect molecules that have an extended expression lifespan in cells to efficiently reprogram somatic cells to the pluripotent state. After 21 days, the non-integrating mc-DNAs disappear and the iPS cells are fully reprogrammed. SBI offers the pre-made, ready-to-transfect 4-in-1 minicircle reprogramming DNA as well as the Human mc-iPS Cell line highlighted in Nature Methods, A nonviral minicircle vector for deriving human iPS cells. Jia F, et al., 2010 Mar;7(3):197-9.
Colonies with morphologies similar to hESC colonies were clearly visible by day 18 after transfection. At day 26–28 after transfection, GFP-negative mc-iPSC colonies were individually picked for further expansion and analysis. The GFP signal decreases over time correlating with the disappearance of the minicricle DNA with simultaneous increase of the endogenous pluripotency marker expression. The bar graph qPCR expression data is adapted from Jia F, et al., 2010.
* SBI is an Authorized and Fully Licensed provider of the Minicircle Reprogramming DNA Kit and the mc-iPS Cell Line.
Retrovirus Features and Benefits
Standard Retroviral-based protein expression system
OSKM Pluripotency factors expressed
Mouse iPS cell colony formation.
The mouse iPS cells were reprogrammed from fibroblasts by using Retroviruses for Oct4, Sox2, cMyc and Klf4. The cells were validated for pluripotent stem cell markers Nanog and SSEA-1 staining. The video shows Mouse iPS cells that were seeded on the glass bottom dish with MEF feeder cells and visualized through a 20X objective lens. The images were taken every 15 minutes for 72 hours at 800x600 resolution with an 1/180 second exposure time. Video created using the Nikon BioStation IM Live Cell Recorder
Features and Benefits
Lentiviral-based protein expression system, HIV backbone for high viral titers
Pluripotency factors expressed from constitutive EF1α promoter
Reliable delivery to dividing or non-dividing cells
Monitor positive expressing cells with co-expressed RFP
All proteins are N-terminal HA-tagged
Convenient sorting of transfected or transduced cells
Create stable cell lines
All 6 iPSC factors expression-validated
Transfection Data
Protein expression Western Blot Data
The piPSC constructs can be used in transient transfection experiments (shown above) or can be used to package lentiviral particles. The particles can then be used to transduce any cell type to create stable cell lines permanently expressing the iPSC protein of interest. Examples of VSV-g pseudotyped lentiviral transductions are shown below.
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