人卵母细胞样细胞体内分化模型的建立

赋予抗TNF-α 单链抗体片段 (TNF-scFv) 对炎症组织的特异性，用一段来自人清蛋白 (HSA) 的柔性连接肽在基因水平上连接TNF-scFv和抗B型纤维连接蛋白 (B-FN) 的额外域B (ED-B) 的scFv L19，构建了抗TNF-α/抗ED-B单链双特异抗体BsDb，其中B-FN为炎症组织中特异表达的抗原。BsDb在毕赤酵母中获得了分泌表达，表达产物经鉴定和纯化制备后，进行了功能分析。结果表明，BsDb保留了其亲本抗体TNF-scFv和L19对抗原的免疫反应性，能够同时结合TNF-α和ED-B，并中和TNF-α的生理作用。而且，BsDb对抗原的亲和力及中和能力与大肠杆菌包涵体来源的亲本抗体相比显著增强。在小鼠佐剂型关节炎 (AIA) 模型中，BsDb能选择性地积累和保留于小鼠的炎症关节，并快速从血浆中清除。说明BsDb兼备炎症组织的特异性和正常组织的低毒性，在类风湿关节炎及其他慢性炎症性疾病的治疗上具有较大潜力。

Generation of human oocyte-like cell differentiation in vivo

Oocyte-like cells (OLC) can be generated by stem cells after the induction and differentiation in vitro, and maturated when transplanted in vivo to improve the development potential. Human amniotic fluid stem cells (hAFSC) were cultured for 10 days in porcine follicle fluid (pFF) that was extracted from the medium follicle with high levels of hormones and Bmp 15 protein. After the induction, the cell aggregates showed the germ cell-like cells and produced the germ cell marker oct4, and triggered epigenetic changes with high expression of methylation transferase gene dnmt3b. The cell aggregates were packaged into porcine theca folliculi to form grafts, which were then transplanted into mouse renal capsule. After one month of transplantation, the morphology of OLC from a graft was not only similar to oocytes, but also expressed the germ cells markers (oct4, nanog, stella, ifitm3, dazl, nanos3, bmp15, and gdf9). The results demonstrate that the in vivo differentiation model was useful for OLC development.