Application of induced pluripotent stem cells for cartilage regeneration in CLAWN miniature pig osteochondral replacement model
Application of induced pluripotent stem cells for cartilage regeneration in CLAWN miniature pig osteochondral replacement model
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DOI：
10.1016/j.reth.2018.06.003
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发表时间：
2018-12-01
影响因子：
4.3
通讯作者：
Hoshi, Kazuto
中科院分区：
文献类型：
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作者：
Uto, Sakura;Nishizawa, Satoru;Hoshi, Kazuto
前言：多能干细胞具有一个优势，即它们可以在不降低质量的情况下增殖，但它们有发生肿瘤的风险。在软骨再生医学中，多能干细胞能够以最小的努力被安全地利用是可取的。为了实现这一目标，我们利用大型动物模型，通过细胞分离和水凝胶包埋的方法，研究了诱导多能干细胞(iPS细胞)在最小处理后用于软骨再生的潜在用途。方法：从CLAWN小型猪建立猪iPS样细胞。用最低限度的处理对猪iPS样细胞进行了体外分化试验。骨软骨置换模型在猪胫骨上端前内侧造成骨软骨缺损。将猪iPS样细胞和人iPS细胞经最低限度处理后种植到热压粘结的β-磷酸三钙和聚L-乳酸支架上，移植到缺损处，观察软骨再生和成瘤情况。结果：体外分析表明，最小处理足以削弱猪iPS样细胞的多能性，而在体外不发生向软骨分化的现象。猪iPS样细胞在体外经最小处理后移植到骨软骨替代模型中，可观察到软骨再生，未见肿瘤形成。此外，荧光原位杂交(FISH)显示，再生软骨中的软骨细胞来源于移植的猪iPS样细胞。人iPS细胞移植在免疫抑制处理的小型猪身上也显示了软骨的再生。结论：最低限度处理的iPS细胞将成为软骨再生医学的有用细胞来源。(C)2018年，日本再生医学学会。爱思唯尔B.V.制作和主办。
Introduction: Pluripotent stem cells have an advantage that they can proliferate without reduction of the quality, while they have risk of tumorigenesis. It is desirable that pluripotent stem cells can be utilized safely with minimal effort in cartilage regenerative medicine. To accomplish this, we examined the potential usefulness of induced pluripotent stem cells (iPS cells) after minimal treatment via cell isolation and hydrogel embedding for cartilage regeneration using a large animal model.Methods: Porcine iPS-like cells were established from the CLAWN miniature pig. In vitro differentiation was examined for porcine iPS-like cells with minimal treatment. For the osteochondral replacement model, osteochondral defect was made in the quarters of the anteromedial sides of the proximal tibias in pigs. Porcine iPS-like cells and human iPS cells with minimal treatment were seeded on scaffold made of thermo-compression-bonded beta-TCP and poly-L-lactic acid and transplanted to the defect, and cartilage regeneration and tumorigenesis were evaluated.Results: The in vitro analysis indicated that the minimal treatment was sufficient to weaken the pluripotency of the porcine iPS-like cells, while chondrogenic differentiation did not occur in vitro. When porcine iPS-like cells were transplanted into osteochondral replacement model after minimal treatment in vitro, cartilage regeneration was observed without tumor formation. Additionally, fluorescent in situ hybridization (FISH) indicated that the chondrocytes in the regenerative cartilage originated from transplanted porcine iPS-like cells. Transplantation of human iPS cells also showed the regeneration of cartilage in miniature pigs under immunosuppressive treatment.Conclusion: Minimally-treated iPS cells will be a useful cell source for cartilage regenerative medicine. (C) 2018, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.