The therapeutic approach for neurological disorder by using novel iPS cell

• Main Authors: Yu-Da Lee, 李昱達
• Other Authors: Shih-Ping Liu 劉詩平
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/26002934181569861690

碩士 === 中國醫藥大學 === 基礎醫學研究所碩士班 === 101 === Both of stroke and Parkinson’s disease are neural injury disease and did not have the effective therapeutic methods. Stem cells therapy provides the hope of treatment. Induced pluripotent stem cells have potential for differentiation ability, and overcome the limitations of embryonic stem cells: immune rejection and ethical issues. BP is one of the major compounds of Angelica sinensis, previous research indicated that BP can maintain stem cells self-renewal and pluripotent ability. We made a novel iPS in the absence of viral infection and oncogenic factors, called iPS-OSH. In this research, we used iPS-OSH to differentiation into neural stem cells and transplanted into mice model to evaluate the therapy effect. In differentiation part, immunofluorescence showed that highly expression of neural stem cell markers: Nestin and Tuj1; RT-PCR showed Oct-4 reduced during the differentiation stage, and Pax6 increased during the differentiation stage. Flow analysis determined that 63% p75 positive cells in all population. All data pointed out that our iPS-OSH success differentiation into neural stem cells. In stroke model, we observed behavoral test improved in therapeutic group including locomotor, beam walking and rotarod in days 14 and 21. Immunofluorescence demonstrated that neural differentiation markers colocalized with iPS-OSH derived NSC in vivo. In Parkinson’s model, behavioral test improved in therapy group (BP, cells and cells + BP co-treatment) after days 5, especially in cells + BP group. In long-term (23 days) data, cell numbers of TH positive cells in therapeutic group are higher then MPTP group at SN. Neural differentiation markers colocalized with bisbenzimind in cells transplanted group. This data indicated that iPS-OSH derived NSC could differentiate into neural cells in vivo. In short-term (5 days) data, cell numbers of TH positive cells in therapeutic groups are higher than MPTP group at SN and striatum. Microglia cells (Iba1) increased in injured brain. The Iba1 positive cells overexpressed in MPTP group and reduced in therapeutic group. In addition, we observed cell proliferation (KI67) in therapy group at SVZ. DA concentration significant declined in MPTP group in days 9, the therapy group recovered to the normal level. TUNEL assay showed that apoptosis level reduced in therapy group. Our study gave a possibility therapeutic approach to Parkinson’s disease and stroke by iPS-OSH derived neural stem cells. It will be a great help for clinical development in the future.