Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030, USA.
Department of Gastroenterolgy, Connecticut Children's Medical Center, 282 Washington Street, Hartford, CT 06106, USA.
Biomaterials. 2015 Jul;57:133-41. doi: 10.1016/j.biomaterials.2015.04.004. Epub 2015 Apr 28.
BACKGROUND/PURPOSE: A tissue-engineered esophagus offers an alternative for the treatment of pediatric patients suffering from severe esophageal malformations, caustic injury, and cancer. Additionally, adult patients suffering from carcinoma or trauma would benefit.
Donor rat esophageal tissue was physically and enzymatically digested to isolate epithelial and smooth muscle cells, which were cultured in epithelial cell medium or smooth muscle cell medium and characterized by immunofluorescence. Isolated cells were also seeded onto electrospun synthetic PLGA and PCL/PLGA scaffolds in a physiologic hollow organ bioreactor. After 2 weeks of in vitro culture, tissue-engineered constructs were orthotopically transplanted.
Isolated cells were shown to give rise to epithelial, smooth muscle, and glial cell types. After 14 days in culture, scaffolds supported epithelial, smooth muscle and glial cell phenotypes. Transplanted constructs integrated into the host's native tissue and recipients of the engineered tissue demonstrated normal feeding habits. Characterization after 14 days of implantation revealed that all three cellular phenotypes were present in varying degrees in seeded and unseeded scaffolds.
We demonstrate that isolated cells from native esophagus can be cultured and seeded onto electrospun scaffolds to create esophageal constructs. These constructs have potential translatable application for tissue engineering of human esophageal tissue.
背景/目的:组织工程食管为治疗患有严重食管畸形、腐蚀性损伤和癌症的儿科患者提供了一种替代方法。此外,患有癌症或创伤的成年患者也将受益。
物理和酶消化供体大鼠食管组织以分离上皮和平滑肌细胞,将其培养在上皮细胞培养基或平滑肌细胞培养基中,并通过免疫荧光进行鉴定。分离的细胞也被播种到电纺合成的 PLGA 和 PCL/PLGA 支架上,在生理中空器官生物反应器中进行培养。体外培养 2 周后,将组织工程构建体进行原位移植。
分离的细胞被证明可以产生上皮细胞、平滑肌细胞和神经胶质细胞类型。培养 14 天后,支架支持上皮细胞、平滑肌细胞和神经胶质细胞表型。移植的构建体与宿主的天然组织整合,接受工程组织的受体表现出正常的进食习惯。植入 14 天后的特征分析表明,所有三种细胞表型在播种和未播种的支架中都以不同程度存在。
我们证明了可以从天然食管中分离出细胞,并将其播种到电纺支架上,以创建食管构建体。这些构建体具有将人类食管组织工程化的潜在转化应用。