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Forskolin is a cell-permeable diterpene that directly activates adenylyl cyclase (IC₅₀ = 41 nM), the enzyme that produces cyclic adenosine monophosphate (cAMP), which as a result raises cAMP levels in the cell. cAMP is an important second messenger involved in many signal transduction pathways, including activation of protein kinase A (PKA; Awad et al.; Robbins et al.).
REPROGRAMMING
· Enables chemical reprogramming (without genetic factors) of mouse embryonic fibroblasts to induced pluripotent stem (iPS) cells, in combination with CHIR99021, Tranylcypromine, Valproic Acid, 3-Deazaneplanocin A, and RepSox (Hou et al.).
· Enables NGN2-mediated transdifferentiation of human fibroblasts to cholinergic neurons (Liu et al.).
· Direct lineage reprogramming of fibroblasts to mature neurons, in combination with RepSox, CHIR99021, SP600125, Valproic Acid, Gö6983 and Y-27632 (Hu et al.).
· Direct lineage reprogramming of fibroblasts to mature neurons, in combination with CHIR99021, ISX-9, SB431542, and I-BET151 (Li et al.).
· Converts human embryonic stem (ES) cells in a “naïve” or ground state similar to mouse ES cells, in combination with LIF, FGF2, TGFβ and small molecules PD0325901, CHIR99021, SP600125, and SB203580 (Hanna et al.).
DIFFERENTIATION
· Potentiates neuronal differentiation of rat hippocampal neural progenitor cells (Hsieh et al., Palmer et al.).
Cell Type
Neural Stem and Progenitor Cells, Neurons, Pluripotent Stem Cells
This product is designed for use in the following research area(s) as part
of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we
offer to support each research area.
Small-Molecule-Driven Direct Reprogramming of Mouse Fibroblasts into Functional Neurons.
Li X et al.
Cell stem cell 2015 AUG
Abstract
Recently, direct reprogramming between divergent lineages has been achieved by the introduction of regulatory transcription factors. This approach may provide alternative cell resources for drug discovery and regenerative medicine, but applications could be limited by the genetic manipulation involved. Here, we show that mouse fibroblasts can be directly converted into neuronal cells using only a cocktail of small molecules, with a yield of up to textgreater90% being TUJ1-positive after 16 days of induction. After a further maturation stage, these chemically induced neurons (CiNs) possessed neuron-specific expression patterns, generated action potentials, and formed functional synapses. Mechanistically, we found that a BET family bromodomain inhibitor, I-BET151, disrupted the fibroblast-specific program, while the neurogenesis inducer ISX9 was necessary to activate neuron-specific genes. Overall, our findings provide a proof of principle" for chemically induced direct reprogramming of somatic cell fates across germ layers without genetic manipulation�
Direct Conversion of Normal and Alzheimer's Disease Human Fibroblasts into Neuronal Cells by Small Molecules.
Hu W et al.
Cell stem cell 2015 AUG
Abstract
Neuronal conversion from human fibroblasts can be induced by lineage-specific transcription factors; however, the introduction of ectopic genes limits the therapeutic applications of such induced neurons (iNs). Here, we report that human fibroblasts can be directly converted into neuronal cells by a chemical cocktail of seven small molecules, bypassing a neural progenitor stage. These human chemical-induced neuronal cells (hciNs) resembled hiPSC-derived neurons and human iNs (hiNs) with respect to morphology, gene expression profiles, and electrophysiological properties. This approach was further applied to generate hciNs from familial Alzheimer's disease patients. Taken together, our transgene-free and chemical-only approach for direct reprogramming of human fibroblasts into neurons provides an alternative strategy for modeling neurological diseases and for regenerative medicine.
Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons.
Liu M-L et al.
Nature communications 2013 JAN
Abstract
Cell fate can be reprogrammed by modifying intrinsic and extrinsic cues. Here we show that two small molecules (forskolin and dorsomorphin) enable the transcription factor Neurogenin 2 (NGN2) to convert human fetal lung fibroblasts into cholinergic neurons with high purity (textgreater90%) and efficiency (up to 99% of NGN2-expressing cells). The conversion is direct without passing through a proliferative progenitor state. These human induced cholinergic neurons (hiCN) show mature electrophysiological properties and exhibit motor neuron-like features, including morphology, gene expression and the formation of functional neuromuscular junctions. Inclusion of an additional transcription factor, SOX11, also efficiently converts postnatal and adult skin fibroblasts from healthy and diseased human patients to cholinergic neurons. Taken together, this study identifies a simple and highly efficient strategy for reprogramming human fibroblasts to subtype-specific neurons. These findings offer a unique venue for investigating the molecular mechanisms underlying cellular plasticity and human neurodegenerative diseases.
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