다발성 경화증 연구 | Human iPSC-derived oligodendrocyte-like cells | 고마바이오텍

특징

• Human O4+ 세포는 OPC 유사 형태를 보이다가, 8일 내에 성숙하여 oligodendrocyte 유사 형태를 갖추게 됩니다. 
• Oligodendrocyte 표지 인자인 PDGFRA, PLP1, MBP, CNP, MAG, MYRF를 발현합니다.
  
• Lot 별로 일관된 whole transcriptome 분석 결과를 보입니다.
• Myelination을 조절하는 화합물을 스크리닝 함으로써, 다발성 경화증(MS)에 대한 Drug Development에 사용할 수 있습니다.
  

사양

• Starting material : Human iPSC line
• Karyotype : 46, XY*
• Seeding compatibility : 6, 12 and 24 well plates
• Shipping info : Dry ice
• Donor : Caucasian adult male (skin fibroblast)
• Vial size : >1 x 10⁶ viable cells
• Quality control : Sterility, protein expression (ICC) and gene expression (RT-qPCR)
• Differentiation method : opti-ox cellular reprogramming
• Recommended seeding density : 27,000 cells/cm²
• User storage : LN2 or -150°C
• Format : Cryopreserved cells
• Product use : ioCells are for research use only
• Applications :
• Screening of compounds that modulate myelination
• Phenotypic assays
• Target validation

실험 결과

그림 1. Immunofluorescent staining of the cells at day 1 (upper panel) and day 8 (lower panel) post-revival. At day 1, the cells are positive for the oligodendrocyte-specific marker O4 (red), and the DAPI counterstain (blue). At day 8, ioOligodendrocyte-like cells show an increased complexity and are positive for O4 (red), the myelin basic protein (MBP) (green), and the DAPI counterstain (blue). 100X magnification; scale bar: 100 μm.

그림 2. Upon reprogramming, cells show rapid morphological changes, acquiring an OPC-like morphology by day 1 post-revival. By day 8, cells have matured and display an oligodendrocyte-like morphology. Brightfield images show day 1 and day 8 post-thawing; scale bar: 400 μm.

그림 3. Following reprogramming, the cells downregulate expression of the pluripotency gene OCT4, whilst demonstrating robust expression of relevant oligodendroglial markers, including PDGFRA, PLP1, MBP, CNP, MAG, and MYRF. Gene expression levels assessed by RT-qPCR, data expressed relative to the reference (housekeeping) gene, HMBS. Data represents day 1 and day 8 post-revival samples; n=2 technical replicates.

그림 4. Bulk RNA-sequencing analysis was performed on three different lots of manufactured product at day 1 and day 8 post revival. Principal component analysis (PCA) represents the variance in gene expression between the three different lots of ioOligodendrocyte-like cells. This analysis shows lots clustering very closely which demonstrates high consistency at each given timepoint. This lot-to-lot consistency in every vial gives scientists confidence in their experimental reproducibility. Colours represent the parental non-induced hiPSC cell line and the three lots of ioOligodendrocyte-like cells; shapes represent different timepoints.