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.