概要: The scRNA-seq analysis of 75 day old human cerebral organoids grown at the air-liquid interface (ALI-COs) reveals six well-defined major clusters. Cell-type and maturity markers define a (1) distinct population of deep layer subcortical projection neurons, (2) upper layer intracortical (callosal) projection neurons and intermediate progenitors, (3) ventricular and subventricular zone radial glial cells, (4) more mature upper and deep layer neurons, (5) interneurons and (6) actively dividing cells with intermediate progenitor and radial glia markers. In addition, the gene expression profile of these clusters corresponds with their expected functional characteristics appropriate to their maturity. Altogether, our data reflect a full repertoire of cortical neuronal and progenitor identities corresponding with the stages of cortical development in age-matched fetal brains.
项目整体设计: Single cell RNA-sequencing data was generated using the 10X Genomics Chromium Single Cell 3' Chip and workflow.
H1 and H9 human embryonic stem cell-derived cerebral organoids were generated and grown as slice cultures as described in Giandomenico et al., 2018. Organoids were supplemented with serum-free slice culture medium (SFSCM): Neurobasal (Invitrogen, cat. #21103049), 1:50 (v/v) B27+A (Invitrogen, cat. #17504044), 0.5% (w/v) glucose, 1X (v/v) Glutamax supplemented with Antibiotic-Antimycotic (ThermoFisher, cat. #15240062). The cultures were maintained in SFSCM at 37 ℃ and 5% CO2 with daily media changes.
处理方案:
-
提取方案:
The cell suspension (34μl with a total of 7000 cells) was loaded onto a 10X Genomics Chromium Single Cell 3' Chip with the appropriate amount of Mastermix. RNA extraction was performed following the same 10X Genomics Chromium workflow.
建库方案:
The reverse transcriptase reaction and subsequent amplification was carried out in a C1000 Touch Thermal Cycler (Biorad), with the cDNA being amplified for 12 cycles. Before sequencing, libraries were quality tested using the 2100 Bioanalyzer Instrument (Agilent) and their concentration was measured by qPCR.
测序信息
分子类型:
poly(A)+ RNA
库的片段类型:
PAIRED
库的链类型:
Forward
测序平台:
ILLUMINA
测序仪型号:
Illumina HiSeq 4000
链特异性:
Specific
样本
基本信息:
样本描述:
生物条件:
实验变量:
方案:
测序信息:
质量评估:
数据来源
GEN样本编号
GEN数据集编号
系列编号
项目编号
样本编号
样本名称
生物样本编号
样本访问号
实验访问号
释放时间
提交时间
最后更新时间
物种
种族
族裔
年龄
年龄单位
性别
来源名称
组织
细胞类型
细胞亚型
细胞系
疾病
疾病状态
发育阶段
突变/变异
表型
Condition Detail
生长方案
处理方案
提取方案
建库方案
分子类型
库的片段类型
链特异性
库的链类型
加标(Spike-In)
测序方法
测序平台
测序仪型号
细胞数
测序片段数
碱基数
平均测序片段长度_1
平均测序片段长度_2
唯一比对率
多重比对率
覆盖度
文章
Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output.