概要: We report the expression of selected genes associated with cortical, striatal and olfactory interneurons in hSS-derived Dlxi1/2::eGFP cells before (in hSS) and after migration (in hCS). By fusing hiPSC-derived fusions that were separately patterned to generate either human subpallial spheroids (hSS) or human cortical spheroids (hCS) we are able to model human interneuron migration in vitro. The aim of this experiment is to examine the gene expression of hSS-derived Dlxi1/2::eGFP cells before and after migration.
项目整体设计: Examination of Dlxi1/2::eGFP cells in hSS (not migrated) and Dlxi1/2::eGFP cells in hCS (migrated).
Differentiation of human iPSC into subpallium-like spheroids; 3D cultures were dissociated and FACS was used to isolate eGFP cells
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The single-cell RNA-Seq library preparation was adapted from the original SMART Seq-2 protocol. Single-cells were FACS sorted into a 96-well PCR plate followed by lysis, reverse transcription and PCR amplification according to SMART-seq2 protocol.
建库方案:
After checking the quality control of cDNA library, final libraries were prepared using Nextera XT library prep kit (96 index primers, Illumina) protocol
测序信息
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poly(A)+ RNA
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SINGLE
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ILLUMINA
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Illumina HiSeq 2500
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Unspecific
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文章
Assembly of functionally integrated human forebrain spheroids.
