瑞士苏黎世联邦理工学院Barbara Treutlein、巴塞尔大学J. Gray Camp等研究人员合作实现人类大脑类器官的谱系记录。相关论文于2021年12月30日在线发表在《自然—方法学》杂志上。

研究人员建立了iTracer，一个结合了报告条码和诱导性CRISPR-Cas9瘢痕的谱系记录器，并且与单细胞和空间转录组学兼容。研究人员应用iTracer来探索了大脑类器官发育过程中的克隆性和系谱动态，并确定了命运限制的时间窗口以及原生神经元家族之间的神经发生动力学变化。研究人员还建立了长期的四维光片显微镜来记录大脑器官中的空间系谱，并确认发育中的神经上皮细胞的区域克隆性。研究人员结合基因扰动（iTracer-perturb）来评估了马赛克TSC2突变对大脑类器官发育的影响。

这些数据揭示了在大脑类器官形成过程中，谱系和命运是如何建立的。更广泛地说，这项技术可以适用于任何诱导多能干细胞（iPSC）衍生的培养系统，用于剖析正常或紊乱发育过程中的谱系改变。

据了解，iPSC衍生的器官提供了研究人类器官发育的模型。单细胞转录组学能够高度解决这些系统内细胞状态的描述，然而，需要有方法来直接测量谱系关系。

附：英文原文

Title: Lineage recording in human cerebral organoids

Author: He, Zhisong, Maynard, Ashley, Jain, Akanksha, Gerber, Tobias, Petri, Rebecca, Lin, Hsiu-Chuan, Santel, Malgorzata, Ly, Kevin, Dupr, Jean-Samuel, Sidow, Leila, Sanchis Calleja, Fatima, Jansen, Sophie M. J., Riesenberg, Stephan, Camp, J. Gray, Treutlein, Barbara

Issue&Volume: 2021-12-30

Abstract: Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enable highly resolved descriptions of cell states within these systems; however, approaches are needed to directly measure lineage relationships. Here we establish iTracer, a lineage recorder that combines reporter barcodes with inducible CRISPR–Cas9 scarring and is compatible with single-cell and spatial transcriptomics. We apply iTracer to explore clonality and lineage dynamics during cerebral organoid development and identify a time window of fate restriction as well as variation in neurogenic dynamics between progenitor neuron families. We also establish long-term four-dimensional light-sheet microscopy for spatial lineage recording in cerebral organoids and confirm regional clonality in the developing neuroepithelium. We incorporate gene perturbation (iTracer-perturb) and assess the effect of mosaic TSC2 mutations on cerebral organoid development. Our data shed light on how lineages and fates are established during cerebral organoid formation. More broadly, our techniques can be adapted in any iPSC-derived culture system to dissect lineage alterations during normal or perturbed development.

DOI: 10.1038/s41592-021-01344-8

Source: https://www.nature.com/articles/s41592-021-01344-8