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iPSC Differentiation

DefiniGEN’s core technology platform has focussed on the development of differentiation protocols which increases the success of generating viable, functional and mature hepatocytes from either patient material (generally fibroblasts and PBMCs) or existing iPSC lines. Our scientific team have optimized differentiation protocols for endodermal cell types across hundreds of different donor lines, both wild-type and diseased, and have established a best-in-class methodology for maximizing the chances of successful differentiation.

Variation in the differentiation capacity of iPSC lines is a well-known phenomenon. DefiniGEN’s platform addresses this in a manner that maximizes the probability of generating high quality hepatocytes from a diverse range of lines. This is achieved by undertaking a set of small scale optimization experiments which provide an assessment of how any particular line will develop during the differentiation process. Using this approach multiple parameters are evaluated in parallel which enables us to further enhance our platform, this maximizes the chances of producing healthy mature hepatocytes at an industrial scale from a particular donor (50m-2bn cells).

 


Workflow

Differentiation of disease-specific human iPSC library into human hepatocytes

 

 

iPSC reprogramming or sourcing: Donor material sourced through DefiniGEN’s hospital network or provided by our clients are reprogrammed to iPSC using multiple methodologies

Differentiation: using the OptiDIFF platform

Cryopreservation: and banking of the differentiated cells

Product validation: genotype, phenotype and cell marker analysis

Delivery: of differentiated cryopreserved cells to client with high viability post-thaw

 


References:

  1. Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells. Sampaziotis F, de Brito MC, Geti I, Bertero A, Hannan NR, Vallier L. Nat Protoc. 2017 Apr;12(4):814-827.
  2. Generation of Hepatocytes from Pluripotent Stem Cells for Drug Screening and Developmental Modeling. Gieseck RL 3rd, Vallier L, Hannan NR. Methods Mol Biol. 2015;1250:123-42.
  3. Generation of Distal Airway Epithelium from Multipotent Human Foregut Stem Cells. Hannan NR, Sampaziotis F, Segeritz CP, Hanley NA, Vallier L. Stem Cells Dev. 2015 Jul 15;24(14):1680-90.
  4. Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells. Cho C , Hannan NR, Vallier L et al. Diabetologia. 2012 Dec 55(12):3284-95.
  5. Targeted gene correction of α1-antitrypsin deficiency in induced pluripotent stem cells. Yusa K, Rashid ST, Vallier L et al. Nature. 2011 Oct12;478(7369):391-4.
  6. Modeling inherited metabolic disorders of the liver using human induced pluripotent stem cells. Rashid ST, Lomas DA, Vallier L et al. J Clin Invest. 2010 Sep;120(9):3127-36.

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