Fisher Robert A, Mas Valeria R
Department of Surgery, Transplantation Division, Virginia Commonwealth University, Medical College of Virginia Hospitals, Richmond, Virginia, USA.
Methods Mol Biol. 2009;481:97-105. doi: 10.1007/978-1-59745-201-4_9.
The use of isolated human hepatocyte infusions to treat human disease will require safe, acceptable, reliable, and reproducible measures of engraftment and function of the donor liver cell. Cell transplant for inborn errors of hepatic metabolism can be followed by measuring the specific protein missing from the recipient, expressed by the transplanted unmodified donor hepatocytes expressing the genes in question. This chapter will focus on the clinical techniques successful in identifying the engraftment and function of donor human hepatocytes when no specific identifiable genes are expressed by donor hepatocytes in acute and chronic liver diseases treated by cell infusion. Radiolabeling and dye labeling techniques, DNA typing of HLA class I alleles, soluble class I HLA ELISA, real-time quantitative PCR techniques including short tandem repeats analysis will be detailed and critiqued.
使用分离的人肝细胞输注来治疗人类疾病将需要安全、可接受、可靠且可重复的方法来衡量供体肝细胞的植入情况和功能。对于先天性肝代谢缺陷的细胞移植，可以通过测量受体中缺失的特定蛋白质来跟踪，该蛋白质由表达相关基因的未修饰供体肝细胞表达。本章将重点介绍在细胞输注治疗的急性和慢性肝病中，当供体肝细胞未表达特定可识别基因时，成功识别供体人肝细胞植入情况和功能的临床技术。将详细介绍和评判放射性标记和染料标记技术、HLA I类等位基因的DNA分型、可溶性I类HLA ELISA、包括短串联重复序列分析在内的实时定量PCR技术。
