Figure 1. Human iPSC-Derived Neural Progenitor Cells Exhibit High-Quality Morphology Characteristic of Multipotent Central Nervous System Progenitor Cells
Cryopreserved Human iPSC-Derived Neural Progenitor Cells were thawed and plated onto Corning® Matrigel®-coated plates at 200,000 cells/cm². NPCs were incubated for 24 hours in STEMdiff™ Neural Progenitor Medium at 37℃ and subsequently analyzed by brightfield microscopy. NPCs display the small, teardrop-shaped morphology expected for NPCs. (A) 4X magnification, (B) 10X magnification. iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
Figure 2. Human iPSC-Derived Neural Progenitor Cells Express Characteristic Markers
Human iPSC-Derived Neural Progenitor Cells generated from SCTi003-A iPSCs were thawed, established in culture, and fixed for immunocytochemistry. The NPCs express neural progenitor markers (A) SOX1 and (B) PAX6 with low expression of (C) βIII-TUB. (D) In addition, they display the expected small, teardrop-shaped morphology. (E) The percentage expression of these markers were quantified. Neural progenitor markers PAX6 and SOX1 were found to be expressed in 90% of NPCs, while mature neuronal marker βIII-TUB was expressed in less than 10% of NPCs. Error bars represent standard deviation (n = 2 biological replicates). iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
Figure 3. Human iPSC-Derived Neural Progenitor Cells Can Effectively Differentiate into Forebrain Neurons
Forebrain neurons were generated from Human iPSC-Derived Neural Progenitor Cells using STEMdiff™ Forebrain Neuron Differentiation Medium. NPCs were cultured for six days at 37℃ in STEMdiff™ Forebrain Neuron Differentiation Medium. Cells were subsequently cultured in STEMdiff™ Forebrain Neuron Maturation medium for 14 days. Resulting cells were processed for immunocytochemistry. (A) The resulting forebrain neuron cultures contain a population of cells expressing (B) neuronal identity marker βIII-TUB (magenta), (C) but not astrocyte marker GFAP (green). (D) Nuclei are labeled with DAPI (gray). iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
Figure 4. Human iPSC-Derived Neural Progenitor Cells Can Effectively Differentiate into Midbrain Neurons
Midbrain neurons were generated from Human iPSC-Derived Neural Progenitor Cells using STEMdiff™ Midbrain Neuron Differentiation Medium. NPCs were cultured for six days at 37℃ in STEMdiff™ Midbrain Neuron Differentiation Medium. Cells were subsequently cultured in STEMdiff™ Midbrain Neuron Maturation medium for 14 days. Resulting cells were processed for immunocytochemistry. (A) The resulting midbrain neuron cultures contain a population of cells expressing (B) neuronal identity marker βIII-TUB (red) and (C) dopaminergic neuron marker TH (green), (D) but not astrocyte marker GFAP (magenta). (D) Nuclei are labeled with DAPI (blue). iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
Figure 5. Human iPSC-Derived Neural Progenitor Cells Can Effectively Differentiate into Astrocytes
Astrocytes were generated from Human iPSC-Derived Neural Progenitor Cells using STEMdiff™ Astrocyte Differentiation Medium. NPCs were cultured for 21 days at 37℃ in STEMdiff™ Astrocyte Differentiation Medium. Cells were subsequently cultured in STEMdiff™ Astrocyte Maturation medium for seven days. Resulting cells were processed for immunocytochemistry. (A) The resulting astrocyte cultures contain a population of cells expressing (B) astrocyte marker S100β (green) and (C) GFAP (red), but not neuronal marker DCX (magenta). Nuclei are labeled with DAPI (blue). iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
Figure 6. Human iPSC-Derived Neural Progenitor Cells Can Be Expanded for Multiple Passages
Human iPSC-Derived Neural Progenitor Cells were thawed and maintained in STEMdiff™ Neural Progenitor Medium at 37℃ and passaged roughly once every seven days for a total of five weeks. The average fold increase of NPCs was 2.8 ± 0.5 (mean ± SEM) per passage, demonstrating cells can be effectively expanded.
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These iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) shall not be used or administered in (1) human subjects for human clinical use; (2) animals for veterinary use for therapeutic, diagnostic, or prophylactic purposes or (3) any subject in relation to clinical applications, cell therapy, transplantation, and/or regenerative medicines, without limiting the generality of the foregoing. These iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) may not be used for monetization or commercialization purposes, including without limitation, used to, or with the goal to, perform services or supply products or rights, including in the manufacture of cellular therapies or other therapeutics, for monetary gain or the generation of royalties, revenues, sales or other valuable consideration. For clarity, these iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) may not be used for screening compounds, antibodies, proteins or peptides, except for the purposes of target discovery, target validation, or assay development, provided such activities and the results of such activities are not further used for monetization or commercialization purposes. It may be possible to obtain a further license for the prohibited uses referred to in this Limited Use License. Please contact iPSCrequests@stemcell.com for more details. UNLESS OTHERWISE STATED, PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES. FOR PRODUCT-SPECIFIC COMPLIANCE AND INTENDED USE INFORMATION, REFER TO THE PRODUCT INFORMATION SHEET. GENERAL INFORMATION ON QUALITY AT STEMCELL MAY BE FOUND AT WWW.STEMCELL.COM/COMPLIANCE.