CAJ | 학술논문

背景：胶原水凝胶可为肝细胞生长和组织重建提供良好的基质支持，并且以胶原为基质构建的工程化组织易于合并生长，形成融合组织。目的：尝试将肝细胞/胶原水凝胶复合物打散成小块肝单元，堆积在皮下腔中，提高工程化肝组织厚度。方法：将新鲜分离的大鼠肝细胞与胶原水凝胶复合，构建肝细胞/胶原水凝胶复合物，待固化后将其打散成小块肝单元，以未打散的肝细胞/胶原水凝胶复合物作为对照。取6只SD大鼠，其中3只切除2/3肝脏，诱导肝再生，于双侧腹股沟皮下腔中分别植入打散与未打散的肝细胞/胶原水凝胶复合物；另3只于双侧腹股沟皮下腔中分别植入打散与未打散的肝细胞/胶原水凝胶复合物。植入后7 d取材，采用苏木精-伊红染色、免疫组化染色、印度墨水灌注等方法对工程化肝组织形成情况进行评价。结果与结论：两组移植物均在皮下腔中形成了血管工程化肝组织，但小块肝单元相互融合，形成了大块血管工程化肝组织，肝组织厚度可达4 mm；整块植入的肝移植物只形成小块肝组织，其厚度只有几层细胞。免疫组织化学染色证实，血管工程化肝组织中的肝细胞具有天然肝细胞的特征及功能。肝部分切除实验表明，工程化肝组织具有对肝部分切除再生刺激产生反应的能力。结果表明将肝细胞/胶原水凝胶移植物打散成小块单元，堆积在皮下腔中，可提高工程化肝组织的厚度。
배경：효원수응효가위간세포생장화조직중건제공량호적기질지지，병차이효원위기질구건적공정화조직역우합병생장，형성융합조직。목적：상시장간세포/효원수응효복합물타산성소괴간단원，퇴적재피하강중，제고공정화간조직후도。방법：장신선분리적대서간세포여효원수응효복합，구건간세포/효원수응효복합물，대고화후장기타산성소괴간단원，이미타산적간세포/효원수응효복합물작위대조。취6지SD대서，기중3지절제2/3간장，유도간재생，우쌍측복고구피하강중분별식입타산여미타산적간세포/효원수응효복합물；령3지우쌍측복고구피하강중분별식입타산여미타산적간세포/효원수응효복합물。식입후7 d취재，채용소목정-이홍염색、면역조화염색、인도묵수관주등방법대공정화간조직형성정황진행평개。결과여결론：량조이식물균재피하강중형성료혈관공정화간조직，단소괴간단원상호융합，형성료대괴혈관공정화간조직，간조직후도가체4 mm；정괴식입적간이식물지형성소괴간조직，기후도지유궤층세포。면역조직화학염색증실，혈관공정화간조직중적간세포구유천연간세포적특정급공능。간부분절제실험표명，공정화간조직구유대간부분절제재생자격산생반응적능력。결과표명장간세포/효원수응효이식물타산성소괴단원，퇴적재피하강중，가제고공정화간조직적후도。
BACKGROUND:Colagen hydrogel provides good matrix support for hepatocyte growth and tissue reconstruction, and the colagen-based engineered tissue is easy to merge the growth and form integrated tissue. OBJECTIVE:To improve the thickness of engineered hepatic tissue by dissociating hepatocytes/colagen hydrogel composite into smal hepatic units that accumulate in the subcutaneous cavity. METHODS:Freshly isolated hepatocytes from rats were mixed with colagen hydrogel to establish hepatocytes/colagen hydrogel composite. The hepatocytes/colagen hydrogel composite was dissociated into smal hepatic units after being cured. The undissociated hepatocytes/colagen hydrogel composite was taken as a control. Six Spraque-Dawley rats were enroled. Three of them were subjected to a two-thirds partial hepatectomy to induce liver regeneration. Dissociated and undissociated hepatocytes/colagen hydrogel composites were implanted into the bilateral inguinal subcutaneous cavity. Dissociated and undissociated hepatocytes/colagen hydrogel composites were implanted into the bilateral inguinal subcutaneous cavity of the other three rats. At the 7th day after transplantation, engineered hepatic tissue formation was evaluated using hematoxylin-eosin staining, immunohistochemical staining and India ink perfusion methods. RESULTS AND CONCLUSION:The grafts in these two groups al formed vascular engineered hepatic tissue in the subcutaneous cavity, but after the smal hepatic units merged, a large piece of vascular engineered hepatic tissue formed. The hepatic tissue thickness was up to 4 mm. The whole piece of implanted liver grafts only formed smal pieces of hepatic tissues, with only several layers of cels. Immunohistochemistry staining confirmed that the hepatocytes in vascular engineered hepatic tissue had the characteristics and functions of natural hepatocytes. Partial hepatectomy experiment showed that engineered hepatic tissue had the ability to respond to regenerative stimulus of partial hepatectomy. These results show that dissociating the hepatocytes/colagen hydrogel grafts into smal units that accumlate in the subcutaneous cavity can increase the thickness of the engineered hepatic tissue.