파킨슨 모델 세포 | Human iPSC-derived Parkinson's Disease model | 고마바이오텍

특징

• Isogenic control(ioGlutamatergic Neurons)과 유사하게 뉴런 특이적인 표지인자를 발현합니다.
• TUBB3 and MAP2 : pan-neuronal protein 
• VGLUT2 : glutamatergic neuron-specific transporter
• 11일차부터 structural neuronal network을 형성합니다.
• 파킨슨 병과 관련된 PRKN R275W이 발현됩니다.
• 세포는 바로 플레이팅할 수 있는 상태로 배송됩니다.

그림 1. Immunofluorescent staining on post-revival day 11 demonstrates similar homogenous expression of pan-neuronal proteins TUBB3 and MAP2 (upper panel) and glutamatergic neuron-specific transporter VGLUT2 (lower panel) in ioGlutamatergic Neurons PRKN R275W/R275W compared to the isogenic control. 100X magnification. 

그림 2. ioGlutamatergic Neurons PRKN R275W/R275W mature rapidly and form structural neuronal networks over 11 days, when compared to the isogenic control. Day 1 to 11 post thawing; 100X magnification.

그림 3. Gene expression analysis demonstrates that ioGlutamatergic Neurons PRKN R275W/R275W and the isogenic control (WT) lack the expression of pluripotency markers (NANOG and OCT4) at day 11, whilst robustly expressing pan-neuronal (TUBB3 and SYP) and glutamatergic specific (VGLUT1 and VGLUT2) markers, as well as the glutamate receptor GRIA4. Gene expression levels were assessed by RT-qPCR (data expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.

그림 4. Gene expression analysis demonstrates that ioGlutamatergic Neurons PRKN R275W/R275W and the isogenic control (WT) express the PRKN gene encoding the Parkin protein. Gene expression levels were assessed by RT-qPCR (data expressed relative to the parental hiPSC control (iPSC Control), normalised to HMBS). Data represents day 11 post-revival samples.

그림 5. ioGlutamatergic Neurons PRKN R275W/R275W are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: Induction, which is carried out at bit.bio (Phase 0), Stabilisation for 4 days (Phase 1), and Maintenance (Phase 2) during which the ioGlutamatergic Neurons PRKN R275W/R275W mature. Phases 1 and 2 after revival of cells are carried out by the customer.

사양

• Starting material : Human iPSC line
• Karyotype : Normal (46, XY)
• Seeding compatibility : 6, 12, 24, 48, 96 & 384 well plates
• Shipping info : Dry ice
• Donor : Caucasian adult male (skin fibroblast)
• Vial size Small : >1 x 10⁶ viable cells
• Quality control : Sterility, protein expression (ICC), gene expression (RT-qPCR) and genotype validation (Sanger sequencing)
• Differentiation method : opti-ox cellular reprogramming
• Recommended seeding density : 30,000 cells/cm²
• User storage : LN2 or -150°C
• Format : Cryopreserved cells
• Genetic modification : 
• Heterozygous R275W missense mutation in the PRKN gene
• Homozygous R275W missense mutation in the PRKN gene
  
• Applications : Parkinson's disease research, Drug discovery and development, Disease modelling
• Product use : ioCells are for research use only

* 사용법은 User Manual을 확인하세요.