Creative Biolabs is dedicated to providing several viable and cost-effective methods for pluripotent stem cells (iPSC) reprogramming. We employ advanced iPSC reprogramming factor delivery strategy by virus, iPSC reprogramming factor delivery by episomal vectors, as well as other iPSC reprogramming methods (mRNA, protein) to help you obtaining the desired iPSC.
Typically, during human disease modeling and cell-based therapies, the capacity to reprogram somatic cells to induce iPSCs has provided great value. Nowadays, iPSC derivatives have already reached the stage of clinical testing in human patients. Using different techniques to produce induced iPSCs needs that scientists choose the suitable reprogramming method for generating iPSCs, thus the resulting iPSCs enable to be transitioned towards clinical applications effectively. Generally steps in a reprogramming experiment include tissue selection, proceeding through iPSC generation, possible transgene excision to produce iPSC cells that are ready for use in a translational setting.
Fig 1. Schematic workflow of iPSC Reprogramming.
Episomal Vectors for iPSC reprogramming are vectors developed to offer the preeminent system for performing transgene-free and virus-free induced iPSCs in a feeder-free environment. Until now, a great number of different somatic cell types have been successfully reprogrammed by using episomal iPSC reprogramming vectors. This advanced system possesses several advantages:
It has proven that direct expression of reprogramming factors as proteins also enable to generate footprint-free iPSCs. Therefore, this approach may be another good choice for the generation of iPSCs appropriate for studies in translational medicine.
Fig 2. Schematic diagram of the generation of human iPSCs from somatic cells of a healthy or diseased individual.
For iPSCs creation, expression of reprogramming factors as mRNAs is also an available zero-footprint technology. However, the strong immunogenic response elicited in cells by the introduction of synthetic mRNA became a bid problem to reprogramming with mRNA. Scientists have solved this problem through modified the RNA bases by substituting 5-methylcytidine for cytidine and pseudouridine for uridine, as well as added the interferon inhibitor B18R into a cell culture media. These modification extremely reduced cell death which derived from the strong antiviral response that is formed when cells come into contact with mRNA containing standard ribonuclease bases in a common cell culture medium.
Minicircle vectors are circularized vectors and their plasmid backbone has been released leaving only the eukaryotic promoter and cDNA(s) that are to be expressed. A minicircle vector composed of Lin28, GFP, Nanog, Sox2, and Oct4, and enables to be used to reprogram human adipose stem cells. Usually, the reprogram steps contain three transfections of the minicircle vector: an initial electroporation, followed by sorting of GFP+ cells, then two lipid-based transfections. With professional scientists devoted themselves in iPSC reprogramming, Creative Biolabs is dedicated to providing the first class iPSC reprogramming services for our customers. Our services will contribute greatly to the success of your iPSC programs. Please contact us for more information and a detailed quote.
References