利用生物脱细胞大鼠肝支架和肝细胞制备生物工程器官。

IF 1.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Organogenesis Pub Date : 2020-04-02 Epub Date: 2020-05-03 DOI:10.1080/15476278.2020.1742534
Tanya Debnath, Chandra Shekar Mallarpu, Lakshmi Kiran Chelluri
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引用次数: 14

摘要

随着对移植器官需求的不断增长,需要开发能够满足器官结构和生理功能的替代品。组织脱细胞和再细胞化有助于保持三维完整性、生化组成、组织超结构和力学行为,使其在功能上适合器官移植。在此,我们尝试在具有翻译潜力的小动物模型(Wistar大鼠)中重建功能性肝移植物。采用0.1%十二烷基硫酸钠(SDS)软法进行脱细胞,原代肝细胞作为潜在的细胞源进行再细胞化。用组织学、DNA含量、超微结构分析对脱细胞过程进行评价和确认。用大鼠肝细胞重新植入支架,并通过其代谢功能和基因表达评估其生物相容性。细胞外基质(ECM)的结构成分(层粘连蛋白、I型胶原蛋白、网状蛋白)被保留,肝细胞特异性蛋白如CK-18、甲胎蛋白、白蛋白在再细胞化支架中表达。从白蛋白和尿素的产生可以看出重建支架的功能和代谢活性。细胞角蛋白-19 (CK-19)、葡萄糖6-磷酸酶(G6P)、白蛋白、γ谷氨酰转移酶(GGT)基因的表达清楚地证实了繁殖过程后的翻译信号。我们的研究清楚地表明,天然的大鼠肝脏细胞外基质可以作为一种支架,用于整个器官再生的有效再细胞化。
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Development of Bioengineered Organ Using Biological Acellular Rat Liver Scaffold and Hepatocytes.

The increasing demand for organs for transplantation necessitates the development of substitutes to meet the structural and physiological functions. Tissue decellularization and recellularization aids in retaining the three-dimensional integrity, biochemical composition, tissue ultra-structure, and mechanical behavior, which makes them functionally suitable for organ transplantation. Herein, we attempted to rebuild functional liver grafts in small animal model (Wistar rat) with a potential of translation. A soft approach was adopted using 0.1% SDS (Sodium Dodecyl Sulfate) for decellularization and primary hepatocytes were used as a potential cell source for recellularization. The decellularization process was evaluated and confirmed using histology, DNA content, ultra-structure analysis. The resultant scaffold was re-seeded with the rat hepatocytes and their biocompatibility was assessed by its metabolic functions and gene expression. The structural components of the Extracellular matrix (ECM) (Laminins, Collagen type I, Reticulins) were conserved and the liver cell-specific proteins like CK-18, alpha-fetoprotein, albumin were expressed in the recellularized scaffold. The functionality and metabolic activity of the repopulated scaffold were evident from the albumin and urea production. Expression of Cytokeratin-19 (CK-19), Glucose 6-Phosphatase (G6P), Albumin, Gamma Glutamyl Transferase (GGT) genes has distinctly confirmed the translational signals after the repopulation process. Our study clearly elucidates that the native extracellular matrix of rat liver can be utilized as a scaffold for effective recellularization for whole organ regeneration.

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来源期刊
Organogenesis
Organogenesis BIOCHEMISTRY & MOLECULAR BIOLOGY-DEVELOPMENTAL BIOLOGY
CiteScore
4.10
自引率
4.30%
发文量
6
审稿时长
>12 weeks
期刊介绍: Organogenesis is a peer-reviewed journal, available in print and online, that publishes significant advances on all aspects of organ development. The journal covers organogenesis in all multi-cellular organisms and also includes research into tissue engineering, artificial organs and organ substitutes. The overriding criteria for publication in Organogenesis are originality, scientific merit and general interest. The audience of the journal consists primarily of researchers and advanced students of anatomy, developmental biology and tissue engineering. The emphasis of the journal is on experimental papers (full-length and brief communications), but it will also publish reviews, hypotheses and commentaries. The Editors encourage the submission of addenda, which are essentially auto-commentaries on significant research recently published elsewhere with additional insights, new interpretations or speculations on a relevant topic. If you have interesting data or an original hypothesis about organ development or artificial organs, please send a pre-submission inquiry to the Editor-in-Chief. You will normally receive a reply within days. All manuscripts will be subjected to peer review, and accepted manuscripts will be posted to the electronic site of the journal immediately and will appear in print at the earliest opportunity thereafter.
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