OPTIMIZATION OF HUMAN FETAL PANCREAS FOR TRANSPLANTATION

人胎胰腺(HFP)具有独特的生长能力 以及移植给糖尿病受体后的分化。 我们资助申请的目的是确定参数 采购、加工、储存、人类白细胞抗原分型和 HFP的细菌和病毒污染检测。我们已经实现了 这些目标并制定了获得高质量的协议 功能良好的组织。我们的研究推动了HFP领域的发展 移植到可以开始临床试验的程度。 限制HFP成功潜力的三个问题仍然存在 临床移植：1)HFP需要三个月以下的时间 在获得葡萄糖反应性之前进行移植 什么时候不可能监测移植排斥反应，2)HFP有一个 组织肿块小，可能需要20个或更多的胰腺才能成功 移植，以及3)HFP移植既要接受同种异体移植 和自身免疫反应。我们的长期目标是取得成功 HFP移植。因此，本申请的目标是： 1)描述同种异体移植物和自身免疫对HFP和 HFP组织的细胞亚群。我们将使用混合淋巴细胞胰岛细胞 培养和特异性淋巴耗竭以确定1)贡献 细胞和细胞因子的反应以及2)直接和 间接抗原提呈导致HFP移植物丢失。我们将使用这个 设计临床相关免疫抑制策略的知识 治疗，2)促进HFP葡萄糖分化 响应性。我们将使用短期培养和腺病毒介导法 特定生长因子和转录因子的基因转移加速 HFP的分化和成熟，以及3)表现出潜在的 瞬时转化HFPβ细胞提高体外培养的HFPβ细胞质量 HFP与SV40-T抗原结合，同时维持β细胞功能。我们会 使用感染了复制的β细胞富集型HFP亚群 表达SV40-T抗原的缺陷腺病毒证实 有可能在不丧失HFP功能的情况下增加β细胞质量。这个 这些研究的结果将提供足够的HFP组织来源 对葡萄糖挑战做出反应的临床移植和 需要最低限度的免疫抑制治疗。

Human fetal pancreas (HFP) has the unique capacity for further growth and differentiation following transplantation to a diabetic recipient. The objective of our funded application was to determine the parameters required for the procurement, processing, storage, HLA typing, and testing for bacterial and viral contamination of HFP. We have achieved these goals and have established protocols for obtaining high quality tissue with good function. Our studies have advanced the field of HFP transplantation to the point where clinical trials can be, initiated. Three problems remain which limit the potential of HFP for successful clinical transplantation: 1) HFP requires up to three months following transplantation before acquisition of glucose responsiveness during which time it is not possible to monitor graft rejection, 2) HFP has a small mass of tissue and may require 20 or more pancreata for successful transplantation, and 3) HFP transplantation is subject to both allograft and autoimmune responses. Our long-term goal is to achieve successful HFP transplantation. Thus the objectives of this application are to: 1) Characterize both the allograft and autoimmune response to HFP and cellular subsets of HFP tissue. We will use mixed lymphocyte islet cell culture and specific lymphoid depletion to determine 1) the contribution of cellular and cytokine responses and 2) the role of direct and indirect antigen presentation to HFP graft loss. We will use this knowledge to design clinically relevant strategies for immunosuppressive therapy, 2) Accelerate the differentiation of HFP glucose responsiveness. We will use short-term culture and adenoviral-mediated gene transfer of specific growth and transcription factors to accelerate HFP differentiation and maturation, and 3) Demonstrate the potential to increase HFP beta cell mass in vitro with transient transformation of HFP with SV40-T antigen while maintaining beta cell function. We will use beta cell enriched HFP subpopulations infected with replication- defective adenovirus expressing SV40-T antigen to demonstrate the potential to increase beta cell mass without loss of HFP function. The results of these studies will provide a sufficient source of HFP tissue for clinical transplantation which responds to glucose challenge and requires minimal immunosuppressive therapy.