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  • Porcine iPSC Generation: Testing Different Protocols to a Successful Application.

Porcine iPSC Generation: Testing Different Protocols to a Successful Application.

Methods in molecular biology (Clifton, N.J.) (2021-12-01)
Oriol Iborra-Egea, Daina Martínez-Falguera, Santiago Roura, Antoni Bayes-Genis, Ángel Raya, Carolina Gálvez-Montón
ABSTRACT

Stem cell therapy has an unparalleled potential to treat blood cancers, cardiovascular diseases and neurodegenerative conditions, among others. However, stem cell therapeutics must overcome multiple requirements before reaching clinical trials, including large animal safety and efficacy studies. In cardiovascular diseases swine models are the most widely adopted due to its great translational potential to humans. In this chapter, we will describe several protocols to induce iPSC dedifferentiation in swine fibroblasts, as well as conditioning treatments that may help in the reprogramming process.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
SB 431542 hydrate, ≥98% (HPLC), powder
Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Sigma-Aldrich
hBFGF, FGF-Basic, recombinant, expressed in E. coli, suitable for cell culture
Roche
Bovine Serum Albumin, blood typing: suitable, Endonuclease, Exonuclease, Rnases, Proteinases free
Sigma-Aldrich
MEK1/2 Inhibitor III, The MEK1/2 Inhibitor III, also referenced under CAS 391210-10-9, controls the biological activity of MEK1/2. This small molecule/inhibitor is primarily used for Phosphorylation & Dephosphorylation applications.
Sigma-Aldrich
5-Aza-2′-deoxycytidine, ≥97%
Sigma-Aldrich
Puromycin dihydrochloride, Ready Made Solution, from Streptomyces alboniger, 10 mg/mL in H2O, suitable for cell culture
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.