In vivo directed differentiation of pluripotent stem cells for skeletal regeneration

Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Plastic and Reconstructive Surgery Division, Department of Medicine and Radiology, and Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 11/2012; 109(50). DOI: 10.1073/pnas.1218052109
Source: PubMed


Pluripotent cells represent a powerful tool for tissue regeneration, but their clinical utility is limited by their propensity to form teratomas. Little is known about their interaction with the surrounding niche following implantation and how this may be applied to promote survival and functional engraftment. In this study, we evaluated the ability of an osteogenic microniche consisting of a hydroxyapatite-coated, bone morphogenetic protein-2-releasing poly-l-lactic acid scaffold placed within the context of a macroenvironmental skeletal defect to guide in vivo differentiation of both embryonic and induced pluripotent stem cells. In this setting, we found de novo bone formation and participation by implanted cells in skeletal regeneration without the formation of a teratoma. This finding suggests that local cues from both the implanted scaffold/cell micro- and surrounding macroniche may act in concert to promote cellular survival and the in vivo acquisition of a terminal cell fate, thereby allowing for functional engraftment of pluripotent cells into regenerating tissue.

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Available from: Jeong S Hyun, Apr 18, 2015
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    • "Moreover, gelatin-based matrices have been demonstrated to degrade [19] [20] [21] [22] [23] [24] and have been studied extensively as a scaffold for tissue engineering [20] [21] [23] [25]. Studies that have reported osteogenic differentiation of hiPSC often used derivation of MSC or mesoderm-like progenitor cells and their subsequent differentiation into osteoblasts, using osteogenic-inducing soluble factors such as b-glycerophosphate, ascorbic acid 2-phosphate, dexamethasone and/or growth factors such as bone morphogenetic protein-2 (BMP-2) [26] [27] [28] [29] [30] [31] [32]. A recent study by de Peppo et al. [27] employed decellularized bone matrix to create bone tissues from hiPSC-derived mesoderm progenitor cells. "
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    ABSTRACT: Human induced pluripotent stem cells (hiPSCs) are a promising cell source with pluripotency and self-renewal properties. Design of simple and robust biomaterials with an innate ability to induce lineage-specificity of hiPSCs is desirable to realize their applications in regenerative medicine. In this study, we investigated the potential of biomaterials containing calcium phosphate minerals to induce osteogenic differentiation of hiPSCs. hiPSCs cultured using mineralized gelatin methacrylate-based matrices underwent osteogenic differentiation ex vivo, both in two- dimensional (2-D) and three-dimensional (3-D) cultures, in growth medium devoid of any osteogenic-inducing chemical components or growth factors. Our findings that osteogenic differentiation of hiPSCs can be achieved through biomaterial-based cues alone present new avenues for personalized regenerative medicine. Such biomaterials that could not only act as structural scaffolds, but could also provide tissue-specific functions such as directing stem cell differentiation commitment, have great potential in bone tissue engineering.
    Full-text · Article · Dec 2014 · Acta Biomaterialia
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    • "Thus, the present strategy can at least partially recapitulate physiological osteoblast development and will be useful for analyzing osteoblast development using gene-manipulated ESCs in vitro. Because the direct differentiation of mouse iPSCs (miPSCs) and human iPSCs (hiPSCs) into osteogenic cells has been previously reported by Bilousova et al. (2011), Kao et al. (2010), and Levi et al. (2012), we applied the present strategy to 2i-adapted miPSCs established from fibroblasts of mice expressing GFP (CAG-GFP miPSCs) (Okabe et al., 1997) (Figure 3A). Nanog was downregulated throughout the culture compared to day 0. T and Mixl1 were transiently upregulated by mesoderm induction on day 5. "
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    ABSTRACT: Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs) and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively) into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenyl)pyrido[4',3':4,5]thieno[2,3-b]pyridine-2-carboxamide [TH]) under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2), a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.
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    • "Several groups have recently demonstrated that progenitors of the mesenchymal lineages (MPs) can be derived from both human embryonic stem cells (hESCs) and hiPSCs [8,16,18-23] and can be further differentiated toward the osteogenic lineage both in vitro and in vivo [8,18,21,24-26]. We discuss the principal strategies for the derivation of MPs, their characteristics in relation to adult hMSCs, and recent advances in constructing bone substitutes from MPs, based on the TE principles developed with hMSCs. "
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    ABSTRACT: Advances in the fields of stem cell biology, biomaterials, and tissue engineering over the last decades have brought the possibility of constructing tissue substitutes with a broad range of applications in regenerative medicine, disease modeling, and drug discovery. Different types of human stem cells have been used, each presenting a unique set of advantages and limitations with regard to the desired research goals. Whereas adult stem cells are at the frontier of research for tissue and organ regeneration, pluripotent stem cells represent a more challenging cell source for clinical translation. However, with their unlimited growth and wide differentiation potential, pluripotent stem cells represent an unprecedented resource for the construction of advanced human tissue models for biological studies and drug discovery. At the heart of these applications lies the challenge to reproducibly expand, differentiate, and organize stem cells into mature, stable tissue structures. In this review, we focus on the derivation of mesenchymal tissue progenitors from human pluripotent stem cells and the control of their osteogenic differentiation and maturation by modulation of the biophysical culture environment. Similarly to enhancing bone development, the described principles can be applied to the construction of other mesenchymal tissues for basic and applicative studies.
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