Stem Cell Unit, Global Medical Education & Research Foundation , Hyderabad, India.
Transplant Immunology & Stem Cell Unit, Gleneagles Global Hospitals , Hyderabad, India.
Organogenesis. 2020 Apr 2;16(2):61-72. doi: 10.1080/15476278.2020.1742534. Epub 2020 May 3.
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.
为了满足器官移植的结构和生理功能需求,对于替代器官的需求不断增加。组织脱细胞和再细胞化有助于保留三维完整性、生化组成、组织超微结构和机械性能,使其在功能上适合器官移植。在此,我们尝试在小型动物模型(Wistar 大鼠)中重建具有潜在转化能力的功能性肝移植物。采用 0.1%SDS(十二烷基硫酸钠)进行脱细胞处理,采用软处理方法,原代肝细胞作为潜在的细胞来源进行再细胞化。通过组织学、DNA 含量、超微结构分析评估和确认脱细胞过程。将大鼠肝细胞重新接种到所得支架上,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、网状蛋白)得以保留,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明显看出,再填充支架的功能和代谢活性。Cytokeratin-19 (CK-19)、葡萄糖 6-磷酸酶(G6P)、白蛋白、γ-谷氨酰转移酶(GGT)基因的表达在再定植过程后明确证实了转化信号。我们的研究清楚地阐明了大鼠肝的固有细胞外基质可以用作整个器官再生的有效再细胞化支架。