Engineering Functional Islets from Cultured Cells
Engineering Functional Islets from Cultured Cells
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DOI：
10.1089/ten.tea.2008.0459
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发表时间：
2009-11-01
影响因子：
4.1
通讯作者：
Vacanti, Charles A.
中科院分区：
文献类型：
--
作者：
Kodama, Shohta;Kojima, Koji;Vacanti, Charles A.
目的：虽然胰岛移植现在可以在1型糖尿病患者中进行微创移植，但功能丰富的胰岛供体的可用性仍然是临床广泛应用的主要障碍。体外组织工程胰岛细胞在体内植入后能够发挥作用，为解决这个问题提供了一种解决方案。研究设计和方法：将大鼠胰岛酶分离成单细胞悬液，种植到聚乙醇酸(PGA)支架上。在含有表皮生长因子、神经生长因子和胰岛素样生长因子的CMRL 1099培养液中培养5d。然后将PGA和分离的细胞悬浮在含有胰岛素、转移和亚硒酸的热可逆明胶聚合物(TGP)中，在F-12和Dulbecco的改良Eagle‘s培养基中增殖40天。在PGA纤维降解后，用冷温萃取法去除TGP。人工采集组织工程(TE)胰岛，移植到链脲佐菌素(STZ)诱导的糖尿病裸鼠肾被膜下。结果：所有接受TE胰岛治疗的小鼠均从诱导的高血糖状态恢复到正常血糖状态(n=6)。接受治疗的小鼠口服葡萄糖耐量试验正常。对物种特异性C肽的测试可以区分TE大鼠胰岛的外源性胰岛素分泌和裸鼠的内源性分泌。免疫组织化学证实了这些TE内分泌细胞的多系潜能，显示它们能够分泌胰岛素、胰升糖素和生长抑素。结论：通过使用PGA和TGP在体外进行胰岛组织工程的能力，在体内完全发挥作用，使糖尿病诱导的小鼠恢复到正常血糖状态，这对治疗1型糖尿病具有潜在的意义。
Objective: Although pancreatic islet transplantation can now be performed minimally invasively in patients with type 1 diabetes, the availability of functional islet donors remains the chief obstacle to widespread clinical application. Tissue engineering islet cells in vitro that function when implanted in vivo provides a solution to this problem. Research Design and Methods: Rat pancreatic islets were enzymatically dissociated into a single-cell suspension and seeded onto a polyglycolic acid (PGA) scaffold. The cells were cultured in CMRL 1099 medium containing epidermal growth factor, nerve growth factor, and insulin-like growth factor for 5 days. The PGA and isolated cells were then suspended in a thermoreversible gelatin polymer (TGP) with insulin, transferring and selenous acid, in F-12 and Dulbecco's modified Eagle's medium, to proliferate over a 40-day period. After the degradation of the PGA fibers, the TGP was removed using cold temperature extraction. The tissue-engineered (TE) islets were then collected manually and transplanted beneath the kidney capsule of Streptozotocin (STZ)induced diabetic nude mice. Results: All mice that received the TE islets reverted from the induced hyperglycemic state to a state of normoglycemia (n = 6). The treated mice demonstrated normal oral glucose tolerance tests. Testing for the species-specific C-peptide allowed discrimination between the exogenous insulin secretions of the TE rat islets and the endogenous secretions of the nude mice. Immunohistochemistry confirmed the multilineage potential of these TE endocrine cells, showing them capable of secreting insulin, glucagon, and somatostatin. Conclusions: The ability to tissue engineer pancreatic islets in vitro, through use of PGA and TGP, that fully function in vivo to return diabetic-induced mice to state of normoglycemia has potential implications for the treatment type 1 diabetes.