Accessing clinical material from the relevant patient population can prove challenging. If you cannot locate human tissue for reprogramming, we can ethically source fresh clinical material for you through our global procurement network.
|Identification of potential donors from your target population||Short-listing of potential donors and collection of ethically approved, informed consent||Screening of blood samples from donors that match your criteria via Next Generation Sequencing (NGS)||Collection of tissue from screened donors for use in your iPSC projects, including blood urine and fibroblasts|
REPROCELL utilizes the most rapid integration-free reprogramming method on the market: StemRNA™ 3rd Gen reprogramming technology. StemRNA™ is up to 50 times more efficient than any other non-integrative methodology on the market*, allowing you to access the highest quality iPSCs for your research.
REPROCELL’s RNA reprogramming is the optimal method for iPSC establishment and our latest generation technology can be used to transform samples from skin, blood and urine. Our methods use RNA transgenes, rather than DNA or viral vectors, which is rapidly degraded by cells. The resultant iPSCs therefore possess low transgene persistence, increased chromosomal stability and reduced clonal variation. As RNA reprogramming leads to decreased copy number variation (CNV), it also reduces the risk of oncogenesis and abnormal karyology.
Whether you have your own cells that require reprogramming or wish to source tissues from a select patient group, our expert scientists have the resources and expertise to achieve your research goals.
*Compared with Episomal reprogramming methodology Epi5 (0.03 - 0.04%), StemRNA™ 3rd generation is at least 50 times more efficient (2.00 - 4.00%).
CRISPR-Cas9 is the ideal gene-editing system for the establishment of disease-relevant models, as it allows the modification of cells in their pluripotent state.
|Identify healthy or patient donors||Harvest donor tissue and establish iPSCs||Insert or delete your desired mutation using CRISPR-Cas9||Select positive clones, culture, and assess novel compounds|
Gene-editing via CRISPR-Cas9 however is an inefficient process in which only a very small percentage of transfected cells are edited in the desired manner. Therefore, many clones must be sequenced in order to identify those that contain your desired mutation; increasing the time spend optimizing cell culture conditions.
The development of SNIPER† has overcome this obstacle by increasing the number of positive clones detected during the initial screening process. In fact, SNIPER is so accurate it can increase the fraction of positive clones detected 300-fold, reducing the number of cells for sequencing. In collaboration with GenAhead Bio®, REPROCELL can now offer CRISPER-SNIPER as part of our stem cell services.
In our SNIPER projects, REPROCELL can also include an isogeneic control to assess the effect of your genetic mutation on disease phenotype. This not only assists the identification of novel test agents but provides a control line for the assessment of drugs tested in your custom model system.
† Specification of Newly Integrated Position and Exclusion of Random-Integration.
As a pioneering company in the production of iPSC-derived cells, REPROCELL has developed relevant protocols for the differentiation of iPSCs into various cell lineages, including neurocytes, hepatocytes and cardiomyocytes. Our know-how and robust manufacturing processes ensure reproducible product and mature cell types that are electrophysiologically responsive with mature morphologies.
Below, you can find an example of custom neuronal differentiated iPSCs generated in partnership with one of our clients, FANCL. Our differentiation technique provides high purity neurons which can be used to model a range of nervous conditions — from bipolar disorder to Parkinson’s disease (Figure 1).
|Figure 1: For the study of Parkinson’s disease, we offer iPSC-derived dopaminergic (DA) neurons. Dopaminergic neurons were generated at high purity without sorting. Immunostaining was used to identify tyrosine hydroxylase (TH)-positive (green) cells. As TH is a DA neuron marker, these results indicated that the differentiation process was up to 60% efficient without sorting.|
We also have a range of media, solutions, and small molecules for stem cell research. Our custom stem cell services include primary cell isolation, iPSC reprogramming, expansion, characterization, banking and differentiation. REPROCELL has decades of experience in custom cell model development and we engage in proprietary collaborations for developing custom differentiation protocols. We can also use or modify your protocol. If you have a particular need, please ask for a project consultation.
Zara Puckrin, BSc / 29 April 2021
Zara Puckrin, BSc / 17 March 2021
Zara Puckrin, BSc / 9 March 2021
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