通过3D生物打印加载的人肝细胞可生成微型肝脏。
Human hepatocytes loaded in 3D bioprinting generate mini-liver.
作者信息
Zhong Cheng, Xie Hai-Yang, Zhou Lin, Xu Xiao, Zheng Shu-Sen
机构信息
Key Laboratory of Combined Multi-organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
出版信息
Hepatobiliary Pancreat Dis Int. 2016 Oct;15(5):512-518. doi: 10.1016/s1499-3872(16)60119-4.
BACKGROUND
Because of an increasing discrepancy between the number of potential liver graft recipients and the number of organs available, scientists are trying to create artificial liver to mimic normal liver function and therefore, to support the patient's liver when in dysfunction. 3D printing technique meets this purpose. The present study was to test the feasibility of 3D hydrogel scaffolds for liver engineering.
METHODS
We fabricated 3D hydrogel scaffolds with a bioprinter. The biocompatibility of 3D hydrogel scaffolds was tested. Sixty nude mice were randomly divided into four groups, with 15 mice in each group: control, hydrogel, hydrogel with L02 (cell line HL-7702), and hydrogel with hepatocyte growth factor (HGF). Cells were cultured and deposited in scaffolds which were subsequently engrafted into livers after partial hepatectomy and radiation-induced liver damage (RILD). The engrafted tissues were examined after two weeks. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, total bilirubin, CYP1A2, CYP2C9, glutathione S-transferase (a-GST), and UDP-glucuronosyl transferase (UGT-2) were compared among the groups. Hematoxylin-eosin (HE) staining and immunohistochemistry of cKit and cytokeratin 18 (CK18) of engrafted tissues were evaluated. The survival time of the mice was also compared among the four groups.
RESULTS
3D hydrogel scaffolds did not impact the viability of cells. The levels of ALT, AST, albumin, total bilirubin, CYP1A2, CYP2C9, a-GST and UGT-2 were significantly improved in mice engrafted with 3D scaffold loaded with L02 compared with those in control and scaffold only (P<0.05). HE staining showed clear liver tissue and immunohistochemistry of cKit and CK18 were positive in the engrafted tissue. Mice treated with 3D scaffold+L02 cells had longer survival time compared with those in control and scaffold only (P<0.05).
CONCLUSION
3D scaffold has the potential of recreating liver tissue and partial liver functions and can be used in the reconstruction of liver tissues.
背景
由于潜在肝移植受者数量与可用器官数量之间的差距日益增大，科学家们试图制造人工肝脏以模拟正常肝功能，从而在肝功能障碍时支持患者的肝脏。3D打印技术符合这一目的。本研究旨在测试3D水凝胶支架用于肝脏工程的可行性。
方法
我们用生物打印机制造了3D水凝胶支架。测试了3D水凝胶支架的生物相容性。60只裸鼠随机分为四组，每组15只：对照组、水凝胶组、含L02（细胞系HL - 7702）的水凝胶组和含肝细胞生长因子（HGF）的水凝胶组。将细胞培养并沉积在支架中，随后在部分肝切除和辐射诱导的肝损伤（RILD）后将支架植入肝脏。两周后检查植入组织。比较各组丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、白蛋白、总胆红素、细胞色素P450 1A2（CYP1A2）、细胞色素P450 2C9（CYP2C9）、谷胱甘肽S -转移酶（α - GST）和尿苷二磷酸葡萄糖醛酸基转移酶（UGT - 2）的水平。对植入组织进行苏木精 - 伊红（HE）染色以及cKit和细胞角蛋白18（CK18）的免疫组织化学评估。还比较了四组小鼠的生存时间。
结果
3D水凝胶支架不影响细胞活力。与对照组和仅使用支架的组相比，植入含L02的3D支架的小鼠中ALT、AST、白蛋白、总胆红素、CYP1A2、CYP2C9、α - GST和UGT - 2的水平显著改善（P<0.05）。HE染色显示肝脏组织清晰，植入组织中cKit和CK18的免疫组织化学呈阳性。与对照组和仅使用支架的组相比，用3D支架 + L02细胞治疗的小鼠生存时间更长（P<0.05）。
结论
3D支架具有重建肝组织和部分肝功能的潜力，可用于肝组织的重建。
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