Natural Scaffolds for Renal Differentiation of Human Embryonic Stem Cells for Kidney Tissue Engineering.
Natural Scaffolds for Renal Differentiation of Human Embryonic Stem Cells for Kidney Tissue Engineering.
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DOI：
10.1371/journal.pone.0143849
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Tarantal AF
中科院分区：
文献类型：
--
作者：
Batchelder CA;Martinez ML;Tarantal AF
尽管人们热衷于移植功能肾组织的生物工程，但在这项技术的潜力能够在临床环境中实现之前，还有许多障碍。可行的肾脏组织工程策略需要鉴定必要的细胞群、高效的支架和3D培养条件，以发展和支持这个重要器官的独特结构和生理功能。我们先前的研究表明，所有年龄段的恒河猴肾脏去细胞切片提供了一种具有足够结构特性的天然细胞外基质(ECM)，对人类胚胎干细胞(HESC)的迁移和分化具有空间和组织上的影响。为了进一步探索脱细胞天然肾支架在肾脏组织工程中的应用，将多能hESC种植在肾脏ECM的整体或切片上，并与已建立的hESC的分化实验进行比较。定量聚合酶链式反应和免疫组织化学分析结果显示，当hESC在无细胞因子或生长因子刺激的脱细胞支架中培养时，肾脏谱系标志物表达上调，提示ECM在引导肾脏谱系分化中起作用。HESC还与生长因子进行了分化，并比较了将其种植在肾脏ECM或新的生物惰性多糖支架上进一步成熟的情况。随着时间的推移，两种支架上的肾脏谱系标志物逐渐上调，hESC被证明表达肾前体细胞、近端小管、内皮细胞和集合管群体的特征基因。这些发现表明，天然支架增强了肾脏谱系标记物的表达，尤其是与胚胎体培养相比。这些研究的结果表明，一种新型的多糖支架具有帮助确定从hESC分化为肾前体细胞的方案的能力，并促进了组织工程作为功能性肾组织来源的前景。
Despite the enthusiasm for bioengineering of functional renal tissues for transplantation, many obstacles remain before the potential of this technology can be realized in a clinical setting. Viable tissue engineering strategies for the kidney require identification of the necessary cell populations, efficient scaffolds, and the 3D culture conditions to develop and support the unique architecture and physiological function of this vital organ. Our studies have previously demonstrated that decellularized sections of rhesus monkey kidneys of all age groups provide a natural extracellular matrix (ECM) with sufficient structural properties with spatial and organizational influences on human embryonic stem cell (hESC) migration and differentiation. To further explore the use of decellularized natural kidney scaffolds for renal tissue engineering, pluripotent hESC were seeded in whole- or on sections of kidney ECM and cell migration and phenotype compared with the established differentiation assays for hESC. Results of qPCR and immunohistochemical analyses demonstrated upregulation of renal lineage markers when hESC were cultured in decellularized scaffolds without cytokine or growth factor stimulation, suggesting a role for the ECM in directing renal lineage differentiation. hESC were also differentiated with growth factors and compared when seeded on renal ECM or a new biologically inert polysaccharide scaffold for further maturation. Renal lineage markers were progressively upregulated over time on both scaffolds and hESC were shown to express signature genes of renal progenitor, proximal tubule, endothelial, and collecting duct populations. These findings suggest that natural scaffolds enhance expression of renal lineage markers particularly when compared to embryoid body culture. The results of these studies show the capabilities of a novel polysaccharide scaffold to aid in defining a protocol for renal progenitor differentiation from hESC, and advance the promise of tissue engineering as a source of functional kidney tissue.