双标记着丝粒和端粒荧光原位杂交技术可鉴定人、大鼠和小鼠细胞对多物种重组泌尿生殖窦异种移植物的贡献。
Dual-label centromere and telomere FISH identifies human, rat, and mouse cell contribution to Multispecies recombinant urogenital sinus xenografts.
作者信息
Vander Griend Donald J, Konishi Yuko, De Marzo Angelo M, Isaacs John T, Meeker Alan K
机构信息
Chemical Therapeutics Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, Maryland 21231, USA.
出版信息
Prostate. 2009 Oct 1;69(14):1557-64. doi: 10.1002/pros.21001.
BACKGROUND
Recombinant xenografts of human cells growing in immunocompromised rodents are widely used for studying stem cell biology, tumor biology, and epithelial to mesenchyme transitions. Of critical importance is the correct interpretation of the cellular composition of such xenografts.
METHODS
Here we present a rapid and robust method employing protein nucleic acid (PNA) FISH probes to dual-label centromeres and telomeres (Cen/Tel FISH). Such labeling allows unambiguous discrimination between human, mouse, and rat cells in paraffin-embedded tissue sections, providing significant advantages over current methods used to discern human versus rodent cell types.
RESULTS
Using an in vivo prostatic developmental system where rat embryonic urogenital sinus mesenchyme is recombined with human prostate epithelial organoids and grown in an immunocompromised mouse, Cen/Tel FISH documents that all three species contribute to the development of glandular structures.
CONCLUSIONS
The method is an indispensable tool to analyze xenograft/host interactions and prevent misinterpretation of data using tissue recombination approaches.
背景
在免疫缺陷啮齿动物体内生长的人细胞重组异种移植广泛用于研究干细胞生物学、肿瘤生物学以及上皮-间充质转化。正确解读此类异种移植的细胞组成至关重要。
方法
在此，我们介绍一种快速且可靠的方法，该方法采用蛋白质核酸（PNA）荧光原位杂交（FISH）探针来对着丝粒和端粒进行双重标记（着丝粒/端粒FISH）。这种标记能够在石蜡包埋的组织切片中明确区分人、小鼠和大鼠细胞，相较于目前用于辨别人类与啮齿动物细胞类型的方法具有显著优势。
结果
利用一种体内前列腺发育系统，将大鼠胚胎泌尿生殖窦间充质与人前列腺上皮类器官重组，并在免疫缺陷小鼠体内生长，着丝粒/端粒FISH证明所有这三个物种都对腺结构的发育有贡献。
结论
该方法是分析异种移植/宿主相互作用以及防止在使用组织重组方法时对数据产生错误解读的不可或缺的工具。
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