布尔肯捷细胞是小脑皮质中唯一的输出神经元，在小脑相关运动协调和记忆整合中起着核心作用。由于小脑萎缩导致的布尔肯捷细胞丧失或功能紊乱会引起严重的共济失调。本研究使用体内移植方法，在转基因小鼠中研究了人诱导多能干细胞衍生的小脑祖细胞的功能，这些小鼠由于多谷氨酸类毒性而导致布尔肯捷特异死亡。以神经祖细胞丰富的小脑器官样结构（培养42-48天）移植后，4周后存活率超过50％。STEM121+ 移植细胞将它们的过程延伸到深小脑核、上小脑脚和前庭小脑核。移植的细胞主要位于白质区域，不在布尔肯捷细胞层中发现。在小脑皮质的分子层中，MAP2阳性纤维接收到爬行纤维的VgluT2输入。经 γ-分泌酶抑制剂DAPT预处理后，移植的神经祖细胞在宿主小脑中过度生长得到抑制。通过使用更成熟的器官样结构（培养86天）或通过FACS分选获得的KIRREL2阳性细胞进行移植，也可抑制过度增殖。移植细胞表达布尔肯捷细胞标志物、GABA、CALB1和L7，尽管它们没有呈扇形形态。我们试图通过移植到成年小鼠中来改善干细胞衍生的小脑祖细胞的神经整合，但未能成功。本研究结果对小脑退行性疾病的再生医学应用和未来小脑发育提供了新的见解。版权所有© 2023 Elsevier Inc. 发表。

Purkinje cells are the sole output neurons of the cerebellar cortex and play central roles in the integration of cerebellum-related motor coordination and memory. The loss or dysfunction of Purkinje cells due to cerebellar atrophy leads to severe ataxia. Here we used in vivo transplantation to examine the function of human iPS cell-derived cerebellar progenitors in adult transgenic mice in which Purkinje-specific cell death occurs due to cytotoxicity of polyglutamines. Transplantation using cerebellar organoids (42-48 days in culture), which are rich in neural progenitors, showed a viability of >50% 4 weeks after transplantation. STEM121+ grafted cells extended their processes toward the deep cerebellar nuclei, superior cerebellar peduncle, and vestibulocerebellar nuclei. The transplanted cells were mostly located in the white matter, and they were not found in the Purkinje cell layer. MAP2-positive fibers seen in the molecular layer of cerebellar cortex received VgluT2 inputs from climbing fibers. Transplanted neural progenitors overgrew in the host cerebellum but were suppressed by pretreatment with the γ-secretase inhibitor DAPT. Hyperproliferation was also suppressed by transplantation with more differentiated organoids (86 days in culture) or KIRREL2-positive cells purified by FACS sorting. Transplanted cells expressed Purkinje cell markers, GABA, CALB1 and L7, though they did not show fan-shaped morphology. We attempted to improve neuronal integration of stem cell-derived cerebellar progenitors by transplantation into the adult mouse, but this was not successfully achieved. Our findings in the present study contribute to regenerative medical application for cerebellar degeneration and provide new insights into cerebellar development in future.Copyright © 2023. Published by Elsevier Inc.