製品の特長

＊本品は、感覚ニューロン前駆細胞から感覚ニューロンへ成熟させるためのキットです。神経堤細胞から感覚ニューロン前駆細胞へ分化させるには、STEMdiff™ Sensory Neuron Differentiation Kit（ST-100-0341）が必要になります。

STEMdiff™ Sensory Neuron培養系で、ヒトES/iPS細胞由来の機能的な感覚ニューロンを作製

• 高コストな後根神経節外植片ワークフローの排除により、動物の使用量を削減
• STEMdiff™ Sensory Neuron Differentiation Kitで作製した感覚ニューロン前駆細胞を維持及び成熟
• 使い勝手のよい簡易な培地仕様で、多能性幹細胞（PSC）由来の感覚ニューロン作製を合理化
• 神経活動と成熟をサポートするBrainPhys™培地により、生理学的関連性の高い結果を取得

感覚ニューロン分化の流れ

感覚ニューロン前駆細胞から7日で感覚ニューロンを作製します。（その前６日間の）hPSC由来NCCから感覚ニューロン前駆細胞への分化については製品添付文書をご確認ください。

Figure 1. Schematic for the STEMdiff™ Sensory Neuron Culture System Protocol

データ紹介

Figure 2. STEMdiff™ Sensory Neuron Kits Promote Differentiation Across Multiple Embryonic Stem and Induced Pluripotent Stem Cell Lines
NCCs generated from hPSCs in mTeSR™ Plus using the STEMdiff™ Neural Crest Differentiation Kit were differentiated and matured to sensory neurons using the STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. (A) Sensory neurons were generated after hPSC-derived NCCs were cultured with the STEMdiff™ Sensory Neuron Differentiation Kit for 6 days and then the STEMdiff™ Sensory Neuron Maturation Kit for 6 days. The resulting cultures contain a population of cells expressing sensory neuron markers peripherin (green) and BRN3A (red) along with (B) neuronal marker class III β-tubulin (TUJ1, red). (C) Midbrain neuron controls generated with STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits do not have detectable peripherin (green) or BRN3A (red) expression, although they express (D) neuronal marker class III β-tubulin (TUJ1, red). Nuclei are labeled with DAPI (blue). Human ES and iPS cell lines were maintained in either mTeSR™1, TeSR™-E8™, or mTeSR™ Plus and differentiated with STEMdiff™ Neural Crest Differentiation Kit, followed by STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. The percentage expression of (E) BRN3A+ and (F) TUJ1+ cells in the resulting cultures was quantified. This differentiation generated BRN3A+ sensory neurons (25.3% ± 6.9%, mean ± SEM; n=7 cell lines, 3 - 23 replicates per condition) that expressed neuronal marker class III β-tubulin (TUJ1; 90.3% ± 4.1%, mean ± SEM; n=4 cell lines, 3 - 12 replicates per condition). Numbers are % positive over total DAPI in a tiled image. NCCs = neural crest cells; hPSCs = human pluripotent stem cells; ES = embryonic stem; iPS = induced pluripotent stem

Figure 3. SNs Derived Using STEMdiff™ Sensory Neuron Differentiation Kit Develop Spontaneous Neuronal Activity Over Time and Respond to Nociceptive Stimulus
(A) Representative images showing H1 mTeSR™ Plus culture-derived differentiation of NCCs to SNs directly in an MEA plate. NCCs were plated at 2 x 105 cells/cm2 and differentiated as described in Methods. SNs grew directly on top of the recording electrodes. (B) Mean firing rate (MFR) was assessed following SN differentiation. There was a gradual increase in MFR for SNs cultured over time, from 0.018 ± 0.008 Hz on day 7 to 2.3 ± 0.4 Hz on day 33 (n=3; mean ± SEM). (C) MFR response to nociceptive stimulus was assessed for midbrain neuron and SN cultures. Day 21 SN cultures displayed a 27 ± 4-fold (mean ± SEM, n=3) increase in MFR in response to temperature increase, which gradually decreased upon recovery. In contrast, day 50 midbrain neurons that displayed spontaneous neuronal activity at 37°C did not display any fold increase in MFR upon temperature increase. Addition of 100 nM capsaicin to SN cultures on day 21 resulted in a similar fold increase in MFR (25 ± 3-fold increase in MFR; mean ± SEM, n=3).