Saturday, July 21, 2012

Converting Human Pluripotent Stem Cells into Nociceptors

Methods for differentiating induced pluripotent stem cells (iPSCs) into specific cell types are a requirement for converting the "promise of iPSCs" into reality. The knowledge derived from this research can be leveraged for high throughput  Drug Discovery and ultimately the development of therapies. I am excited to highlight results recently published by Dr. Lorenz Studer and his team at Memorial Sloan-Ketterling: Stuart M Chambers, Yuchen Qi, Yvonne Mica, Gabsang Lee, Xin-Jun Zhang, Lei Niu, James Bilsland, Lishuang Cao, Edward Stevens, Paul Whiting, Song-Hai Shi, Lorenz Studer. Combined small-molecule inhibition accelerates developmental timing and converts human pluripotent stem cells into nociceptors. Nature Biotechnology 30, 715–720 (2012) doi:10.1038/nbt.2249.

Abstract: Considerable progress has been made in identifying signaling pathways that direct the differentiation of human pluripotent stem cells (hPSCs) into specialized cell types, including neurons. However, differentiation of hPSCs with extrinsic factors is a slow, step-wise process, mimicking the protracted timing of human development. Using a small-molecule screen, we identified a combination of five small-molecule pathway inhibitors that yield hPSC-derived neurons at >75% efficiency within 10 d of differentiation. The resulting neurons express canonical markers and functional properties of human nociceptors, including tetrodotoxin (TTX)-resistant, SCN10A-dependent sodium currents and response to nociceptive stimuli such as ATP and capsaicin. Neuronal fate acquisition occurs about threefold faster than during in vivo development(1), suggesting that use of small-molecule pathway inhibitors could become a general strategy for accelerating developmental timing in vitro. The quick and high-efficiency derivation of nociceptors offers unprecedented access to this medically relevant cell type for studies of human pain

Figure – LSB3i Differentiation model. Early LSB inhibits trophectoderm, mesendoderm, and non-neural ectoderm cell fates yielding neuroectoderm. CHIR99021, SU5402 and DAPT induce and accelerate neural crest stem cell identity by day 8 and promote rapid differentiation of the neural crest stem cells to nociceptors expressing peptidergic markers by day 10.
Note: Neuromics' TRPV1 Antibody was used as a marker for mature nociceptors.

Check out Supplementary Data for more. I will continue to post links to methods here and @ Neuromics' Stem Cell Research Reagents.

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