STEMCELL Technologies STEMdiff Human iPSC-Derived Microglia
- 研究用
- 新製品
本製品は高品質でReady-to-useの凍結ヒト人工多能性幹細胞（iPSC）由来ミクログリアであり、神経研究を確実に加速させることができます。ミクログリアはSTEMdiff™ Microglia Maturation Kit（商品コード：ST-100-0020）で解凍後、直ちに下流アプリケーションに使用可能です。細胞は高純度（> 70%CD45+/CD11b+, P2RY12+TREM2+）であり、非活性化状態でありながら機能的です。本製品は高度に特性解析されたヒトiPSCコントロール株 SCTi003-A（商品コード：ST-200-0511）を基に血清フリーのシステムで製造されています。解凍後は最大20日間維持可能であり、神経炎症シグナル経路の解析、ヒト神経発生および疾患モデルの構築、共培養、創薬スクリーニング、毒性試験、細胞治療の検証など、さまざまな用途にご利用いただけます。
本製品は研究用製品（RUO）であり、学術・商業研究用としての承認を受けています。血液サンプルは、治験審査委員会（IRB）またはその他の規制当局が承認した同意書およびプロトコルを用いて倫理的に提供されています。
2018/05/14 12:00の製品情報
本製品は研究目的にのみ使用し、人や動物の医療用・臨床診断用・食品用としては使用しないようにご注意ください。
製品の特長
ヒトiPS細胞株SCTi003-Aから分化したミクログリア
- Ready-to-use：高度に特性解析されたヒトiPSCコントロール株由来の凍結ヒトミクログリアで作業時間短縮
- 高品質：非活性化状態で機能的かつ高純度のミクログリア
- 優れた安定性：STEMdiff™ Microglia Maturation Kitを用いて、最大20日間の培養維持が可能
- 生理的関連性：遺伝的バックグラウンドを揃えたニューロン-グリア共培養により、生理的関連性の高い培養が可能
- 高い汎用性：神経発生・疾患モデル、創薬スクリーニング、毒性評価、細胞治療検証など、多様な用途に柔軟に対応可能
ご注意
ソースセルバンクのドナー詳細および細胞品質特性については、以下のSCTi003-Aの情報をご参照ください。
コントロールに最適な凍結ヒトiPS細胞株
SCTi003-Aは、αβT細胞に由来し、VDJ配列の再構成を受けています。核型的に安定であり、3胚葉分化能を示し、未分化細胞マーカーを発現し、非組み込み型技術によりリプログラミングされています。hPSCreg®に登録されており、コミュニティ基準に基づく倫理的・生物学的適合性が保証されます。詳細については、ロット別の検査証明書（CoA）およびiPSC株に関するよくある質問もご参照ください。
データ紹介
Figure 1. Human iPSC-Derived Microglia Exhibit High-Quality Morphology
Cryopreserved Human iPSC-Derived Microglia generated from SCTi003-A iPSCs were thawed and plated onto Matrigel®-coated plates at 1.1 x 10^4 cells/cm² in STEMdiff™ Microglia Maturation Kit. Microglia were incubated at 37°C and subsequently imaged on Days 2, 10 (before replating), and 20 (after replating) by brightfield microscopy at 10X magnification. Microglia display a compact morphology with moderate ramification due to reduced adherence, as expected in a healthy homeostatic microglia monoculture. iPSC = induced pluripotent stem cell.
Figure 2. Human iPSC-Derived Microglia Exhibit High Purity and Express Characteristic Glial Markers
Cryopreserved Human iPSC-Derived Microglia generated from SCTi003-A iPSCs were thawed and cultured using STEMdiff™ Microglia Maturation Kit. Microglia co-expressed CD45 and CD11b and expressed P2RY12 and TREM2 markers, as measured by flow cytometry on Days 0 and 10 post-thaw. The bars show the mean ± standard deviation. Technical replicates (n = 1 - 2) were averaged, and each dot in the graph represents an experimental replicate with different manufacturing lots separated by color. iPSC = induced pluripotent stem cell.
Figure 3. Human iPSC-Derived Microglia Display Cytokine Release in Response to Inflammatory Stimuli
Cryopreserved Human iPSC-Derived Microglia generated from SCTi003-A iPSCs were thawed and plated at 100,000 cells per well in a 24-well plate using STEMdiff™ Microglia Maturation Kit for 1 day. Microglia were treated with water (control) or stimulated with 100 ng/mL LPS for 24 hours. (A) Brightfield microscopy shows the expected microglia morphology, including small cell bodies and ramified processes, under vehicle control conditions. After LPS treatment, activated microglial morphology with larger cell bodies, hypertrophic ramification, ameboid-like or rod-like shapes is observed. (B) The release of pro-inflammatory cytokines (TNFα and IL-6) was measured by Duoset ELISA with 20X dilution of the LPS-treated supernatants and the undiluted control supernatants. Microglia release cytokines in response to LPS treatment, as expected. Technical replicates (n = 2) were averaged, and each dot in the graph represents an experimental replicate with different manufacturing lots separated by color.*** p ≤ 0.001 and **** p ≤ 0.0001. LPS = lipopolysaccharide; iPSC = induced pluripotent stem cell.
Figure 4. Human iPSC-Derived Microglia Are Capable of Phagocytosis
Cryopreserved Human iPSC-Derived Microglia generated from SCTi003-A iPSCs were thawed and cultured using STEMdiff™ Microglia Maturation Kit for 1 day. Phagocytosis was assessed by adding pH-sensitive bioindicator particles (25 µg/mL), and particle uptake was tracked over 24 hours using live-cell imaging. As the particles are phagocytosed, the particles turn red and are concentrated within the cells. Over time, the microglia display an activated ameboid morphology. iPSC = induced pluripotent stem cell.
Figure 5. Human iPSC-Derived Microglia Can Be Tri-Cultured with Neurons and Astrocytes to Model Cell-Cell Interactions In Vitro
Representative immunocytochemistry images display Human iPSC-Derived Microglia tri-cultured with Human iPSC-Derived Astrocytes and Human iPSC-Derived Forebrain Neurons for 7 or 20 days using BrainPhys™ hPSC Neuron Kit supplemented with STEMdiff™ Microglia Supplement 2 (Component #100-0023 of STEMdiff™ Microglia Differentiation Kit). Cells were stained for MAP2 (cyan, neuronal dendrite marker), IBA1 (pink, microglia marker), GFAP (green, astrocyte marker), and DAPI (grey, nuclei). The presence of MAP2-positive neurons, GFAP-positive astrocytes, and IBA1-positive microglia confirms the successful establishment of tri-culture conditions. iPSC = induced pluripotent stem cell.
