Park Thomas I H, Waldvogel Henry J, Montgomery Johanna M, Mee Edward W, Bergin Peter S, Faull Richard L M, Dragunow Mike, Curtis Maurice A
Department of Pharmacology and Clinical Pharmacology, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand.
Centre for Brain Research, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand.
Methods Mol Biol. 2022;2389:125-154. doi: 10.1007/978-1-0716-1783-0_12.
The discovery, in 1998, that the adult human brain contains at least two populations of progenitor cells and that progenitor cells are upregulated in response to a range of degenerative brain diseases has raised hopes for their use in replacing dying brain cells. Since these early findings, the race has been on to understand the biology of progenitor cells in the human brain, and they have now been isolated and studied in many major neurodegenerative diseases. Before these cells can be exploited for cell replacement purposes, it is important to understand how to (1) locate them, (2) label them, (3) determine what receptors they express, (4) isolate them, and (5) examine their electrophysiological properties when differentiated. In this chapter we have described the methods we use for studying progenitor cells in the adult human brain and in particular the tissue processing, immunohistochemistry, autoradiography, progenitor cell culture, and electrophysiology on brain cells. The Neurological Foundation of New Zealand Human Brain Bank has been receiving human tissue for approximately 25 years during which time we have developed a number of unique ways to examine and isolate progenitor cells from resected surgical specimens as well as from postmortem brain tissue. There are ethical and technical considerations that are unique to working with human brain tissue, and these, as well as the processing of this tissue and the culturing of it for the purpose of studying progenitor cells, are the topic of this chapter.
1998年有两项发现：一是成人大脑中至少含有两类祖细胞；二是祖细胞会因一系列退行性脑疾病而上调，这让人们燃起了利用它们来替代死亡脑细胞的希望。自这些早期发现以来，人们一直在竞相了解人脑中祖细胞的生物学特性，目前已经在多种主要神经退行性疾病中对其进行了分离和研究。在能够利用这些细胞进行细胞替代之前，了解如何（1）定位它们、（2）标记它们、（3）确定它们表达何种受体、（4）分离它们以及（5）在分化时检测它们的电生理特性非常重要。在本章中，我们描述了我们用于研究成人大脑中祖细胞的方法，特别是脑组织处理、免疫组织化学、放射自显影、祖细胞培养以及脑细胞电生理学方法。新西兰人类脑库神经基金会接收人类组织已有约25年时间，在此期间我们开发了许多独特的方法，用于从切除的手术标本以及死后脑组织中检测和分离祖细胞。处理人类脑组织有一些独特的伦理和技术考量，本章的主题就是这些考量以及为研究祖细胞而对该组织进行的处理和培养。
