Protocol for isolating viable human central nervous system T cells.
Paper
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The authors' code
Shell · 10 lines · 295 B · MIT
- # usage:
- # extractSJfromGTF.sh in.gtf > out.sj
- #
- # assumes transcript_id in the 12th field of GTF
- #
- awk '$3=="exon" {print $12,$1,$4,$5,$7}' $1 |\
- sort -k1,1V -k2,2V -k3,3n |\
- awk 'BEGIN {OFS="\t"} {if (t==$1) {print $2,e1+1,$3-1,$5}; e1=$4;t=$1 }' |\
- sort -k1,1V -k2,2n -k3,3n -k4,4 | uniq
extractSJfromGTF.sh at commit b1edc12, under MIT · at the source
Overview
- Neuroimmunology Research Group, Netherlands Institute for Neuroscience, Amsterdam 1105 BA, the Netherlands
- Department of Immunology, MS Center ErasMS, Erasmus MC, University Medical Center Rotterdam, Rotterdam 3015 GD, the Netherlands
- Department of Neurology, MS Center ErasMS, Erasmus MC, University Medical Center Rotterdam, Rotterdam 3015 GD, the Netherlands
- Swammerdam Institute for Life Sciences, Center for Neuroscience, University of Amsterdam, Amsterdam 1098 XH, the Netherlands
- Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam 1105 BA, the Netherlands
- Department of Experimental Immunology, Amsterdam Institute for Immunology and Infectious Diseases, Amsterdam University Medical Center, Amsterdam 1105 AZ, the Netherlands
Abstract
Brain-resident T cells act as sentinels, monitoring and supporting immune surveillance and homeostasis. Here, we present a rapid protocol for isolating viable T cells from post-mortem human brain tissue. We describe the process of extracting cells from multiple CNS compartments—including choroid plexus, leptomeninges, dura mater, cerebrospinal fluid, and parenchyma—as well as matched peripheral blood. We detail steps for achieving this through mechanical and enzymatic tissue dissociation, followed by density gradient centrifugation to isolate mononuclear cells for downstream applications.
For complete details on the use and execution of this protocol, please refer to Hsiao et al.1
Reproduced under the paper's license (CC BY), from the paper cited above.
Repository
Its files are read in the Code ↔ Paper reader above.
alexdobin/STAR
b1edc1208d91a53bf40ebae8669f71d50b994851, 25 January 2024Availability: 2 checks, the latest on 28 October 2026: the link answers
- 28 October 2026: the link answers
- 27 September 2026: the link answers
331 files
- extras/
scripts/ , Shell, 10 linesextractSJfromGTF.sh - extras/
scripts/ , MATLAB, 45 linessjMotif.m - source/
AlignVsTranscript.h , C/C++, 9 lines - source/
BAMbinSortByCoordinate.c , C++, 81 linespp - source/
BAMbinSortByCoordinate.h , C/C++, 12 lines - source/
BAMbinSortUnmapped.cpp , C++, 91 lines - source/
BAMbinSortUnmapped.h , C/C++, 12 lines - source/
BAMfunctions.cpp , C++, 194 lines - source/
BAMfunctions.h , C/C++, 79 lines - source/
BAMoutput.cpp , C++, 186 lines - source/
BAMoutput.h , C/C++, 37 lines - source/
Chain.cpp , C++, 126 lines - source/
Chain.h , C/C++, 30 lines - source/
ChimericAlign.cpp , C++, 32 lines - source/
ChimericAlign.h , C/C++, 41 lines - source/
ChimericAlign_chimericBA , C++, 105 linesMoutput.cpp - source/
ChimericAlign_chimericJu , C++, 23 linesnctionOutput.cpp - source/
ChimericAlign_chimericSt , C++, 181 linesitching.cpp - source/
ChimericDetection.cpp , C++, 6 lines - source/
ChimericDetection.h , C/C++, 29 lines - source/
ChimericDetection_chimer , C++, 138 linesicDetectionMult.cpp - source/
ChimericSegment.cpp , C++, 32 lines - source/
ChimericSegment.h , C/C++, 23 lines - source/
ChimericTranscript.h , C/C++, 19 lines - source/
ClipCR4.cpp , C++, 109 lines - source/
ClipCR4.h , C/C++, 41 lines - source/
ClipMate.h , C/C++, 34 lines - source/
ClipMate_clip.cpp , C++, 78 lines - source/
ClipMate_clipChunk.cpp , C++, 62 lines - source/
ClipMate_initialize.cpp , C++, 32 lines - source/
ErrorWarning.cpp , C++, 37 lines - source/
ErrorWarning.h , C/C++, 9 lines - source/
GTF.cpp , C++, 171 lines - source/
GTF.h , C/C++, 36 lines - source/
GTF_superTranscript.cpp , C++, 257 lines - source/
GTF_transcriptGeneSJ.cpp , C++, 183 lines - source/
Genome.cpp , C++, 234 lines - source/
Genome.h , C/C++, 107 lines - source/
Genome_genomeGenerate.cp , C++, 438 linesp - source/
Genome_genomeLoad.cpp , C++, 521 lines - source/
Genome_genomeOutLoad.cpp , C++, 58 lines - source/
Genome_insertSequences.c , C++, 33 linespp - source/
Genome_transformGenome.c , C++, 324 linespp - source/
GlobalVariables.cpp , C++, 4 lines - source/
GlobalVariables.h , C/C++, 9 lines - source/
InOutStreams.cpp , C++, 44 lines - source/
InOutStreams.h , C/C++, 23 lines - source/
IncludeDefine.h , C/C++, 255 lines - source/
OutSJ.cpp , C++, 123 lines - source/
OutSJ.h , C/C++, 59 lines - source/
PackedArray.cpp , C++, 43 lines - source/
PackedArray.h , C/C++, 34 lines - source/
ParameterInfo.h , C/C++, 112 lines - source/
Parameters.cpp , C++, 1,269 lines - source/
Parameters.h , C/C++, 374 lines - source/
ParametersChimeric.h , C/C++, 41 lines - source/
ParametersChimeric_initi , C++, 118 linesalize.cpp - source/
ParametersClip.h , C/C++, 37 lines - source/
ParametersClip_initializ , C++, 107 linese.cpp - source/
ParametersGenome.cpp , C++, 59 lines - source/
ParametersGenome.h , C/C++, 63 lines - source/
ParametersSolo.cpp , C++, 721 lines - source/
ParametersSolo.h , C/C++, 220 lines - source/
Parameters_closeReadsFil , C++, 12 lineses.cpp - source/
Parameters_openReadsFile , C++, 111 liness.cpp - source/
Parameters_readFilesInit , C++, 167 lines.cpp - source/
Parameters_readSAMheader , C++, 44 lines.cpp - source/
Parameters_samAttributes , C++, 263 lines.cpp - source/
Quantifications.cpp , C++, 37 lines - source/
Quantifications.h , C/C++, 22 lines - source/
ReadAlign.cpp , C++, 125 lines - source/
ReadAlign.h , C/C++, 239 lines - source/
ReadAlignChunk.cpp , C++, 170 lines - source/
ReadAlignChunk.h , C/C++, 49 lines - source/
ReadAlignChunk_mapChunk. , C++, 128 linescpp - source/
ReadAlignChunk_processCh , C++, 302 linesunks.cpp - source/
ReadAlign_CIGAR.cpp , C++, 62 lines - source/
ReadAlign_alignBAM.cpp , C++, 614 lines - source/
ReadAlign_assignAlignToW , C++, 130 linesindow.cpp - source/
ReadAlign_calcCIGAR.cpp , C++, 58 lines - source/
ReadAlign_chimericDetect , C++, 57 linesion.cpp - source/
ReadAlign_chimericDetect , C++, 312 linesionOld.cpp - source/
ReadAlign_chimericDetect , C++, 74 linesionOldOutput.cpp - source/
ReadAlign_chimericDetect , C++, 39 linesionPEmerged.cpp - source/
ReadAlign_createExtendWi , C++, 85 linesndowsWithAlign.cpp - source/
ReadAlign_mapOneRead.cpp , C++, 118 lines - source/
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ReadAlign_oneRead.cpp , C++, 124 lines - source/
ReadAlign_outputAlignmen , C++, 325 linests.cpp - source/
ReadAlign_outputSpliceGr , C++, 164 linesaphSAM.cpp - source/
ReadAlign_outputTranscri , C++, 65 linesptCIGARp.cpp - source/
ReadAlign_outputTranscri , C++, 359 linesptSAM.cpp - source/
ReadAlign_outputTranscri , C++, 56 linesptSJ.cpp - source/
ReadAlign_outputVariatio , C++, 13 linesn.cpp - source/
ReadAlign_peOverlapMerge , C++, 368 linesMap.cpp - source/
ReadAlign_quantTranscrip , C++, 91 linestome.cpp - source/
ReadAlign_stitchPieces.c , C++, 350 linespp - source/
ReadAlign_stitchWindowSe , C++, 278 lineseds.cpp - source/
ReadAlign_storeAligns.cp , C++, 160 linesp - source/
ReadAlign_transformGenom , C++, 74 linese.cpp - source/
ReadAlign_waspMap.cpp , C++, 128 lines - source/
ReadAnnotations.h , C/C++, 47 lines - source/
STAR.cpp , C++, 313 lines - source/
SequenceFuns.cpp , C++, 445 lines - source/
SequenceFuns.h , C/C++, 32 lines - source/
SharedMemory.cpp , C++, 291 lines - source/
SharedMemory.h , C/C++, 182 lines - source/
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SjdbClass.h , C/C++, 17 lines - source/
Solo.cpp , C++, 91 lines - source/
Solo.h , C/C++, 30 lines - source/
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SoloBarcode.h , C/C++, 38 lines - source/
SoloBarcode_extractBarco , C++, 34 linesde.cpp - source/
SoloCommon.h , C/C++, 80 lines - source/
SoloFeature.cpp , C++, 65 lines - source/
SoloFeature.h , C/C++, 110 lines - source/
SoloFeatureTypes.h , C/C++, 12 lines - source/
SoloFeature_addBAMtags.c , C++, 26 linespp - source/
SoloFeature_cellFilterin , C++, 131 linesg.cpp - source/
SoloFeature_collapseUMI_ , C++, 172 linesGraph.cpp - source/
SoloFeature_collapseUMIa , C++, 658 linesll.cpp - source/
SoloFeature_countCBgeneU , C++, 112 linesMI.cpp - source/
SoloFeature_countSmartSe , C++, 159 linesq.cpp - source/
SoloFeature_countVelocyt , C++, 166 lineso.cpp - source/
SoloFeature_emptyDrops_C , C++, 231 linesR.cpp - source/
SoloFeature_loadRawMatri , C++, 141 linesx.cpp - source/
SoloFeature_outputResult , C++, 241 liness.cpp - source/
SoloFeature_processRecor , C++, 87 linesds.cpp - source/
SoloFeature_quantTranscr , C++, 327 linesipt.cpp - source/
SoloFeature_redistribute , C++, 80 linesReadsByCB.cpp - source/
SoloFeature_statsOutput. , C++, 123 linescpp - source/
SoloFeature_sumThreads.c , C++, 101 linespp - source/
SoloFilteredCells.h , C/C++, 33 lines - source/
SoloRead.cpp , C++, 22 lines - source/
SoloRead.h , C/C++, 23 lines - source/
SoloReadBarcode.cpp , C++, 51 lines - source/
SoloReadBarcode.h , C/C++, 46 lines - source/
SoloReadBarcodeStats.h , C/C++, 27 lines - source/
SoloReadBarcode_getCBand , C++, 429 linesUMI.cpp - source/
SoloReadFeature.cpp , C++, 63 lines - source/
SoloReadFeature.h , C/C++, 50 lines - source/
SoloReadFeatureStats.h , C/C++, 49 lines - source/
SoloReadFeature_inputRec , C++, 172 linesords.cpp - source/
SoloReadFeature_record.c , C++, 276 linespp - source/
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SpliceGraph.cpp , C++, 33 lines - source/
SpliceGraph.h , C/C++, 58 lines - source/
SpliceGraph_findSuperTr. , C++, 186 linescpp - source/
SpliceGraph_swScoreSplic , C++, 249 linesed.cpp - source/
SpliceGraph_swTraceBack. , C++, 19 linescpp - source/
Stats.cpp , C++, 156 lines - source/
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SuffixArrayFuns.cpp , C++, 410 lines - source/
SuffixArrayFuns.h , C/C++, 18 lines - source/
SuperTranscriptome.cpp , C++, 92 lines - source/
SuperTranscriptome.h , C/C++, 42 lines - source/
ThreadControl.cpp , C++, 7 lines - source/
ThreadControl.h , C/C++, 29 lines - source/
TimeFunctions.cpp , C++, 20 lines - source/
TimeFunctions.h , C/C++, 9 lines - source/
Transcript.cpp , C++, 59 lines - source/
Transcript.h , C/C++, 83 lines - source/
Transcript_alignScore.cp , C++, 60 linesp - source/
Transcript_convertGenome , C++, 161 linesCigar.cpp - source/
Transcript_generateCigar , C++, 63 linesP.cpp - source/
Transcript_transformGeno , C++, 167 linesme.cpp - source/
Transcript_variationAdju , C++, 74 linesst.cpp - source/
Transcript_variationOutp , C++, 6 linesut.cpp - source/
Transcriptome.cpp , C++, 190 lines - source/
Transcriptome.h , C/C++, 63 lines - source/
Transcriptome_alignExonO , C++, 278 linesverlap.cpp - source/
Transcriptome_classifyAl , C++, 267 linesign.cpp - source/
Transcriptome_geneCounts , C++, 63 linesAddAlign.cpp - source/
Transcriptome_geneFullAl , C++, 56 linesignOverlap.cpp - source/
Transcriptome_geneFullAl , C++, 43 linesignOverlap_ExonOverIntro n.cpp - source/
Transcriptome_quantAlign , C++, 114 lines.cpp - source/
Variation.cpp , C++, 157 lines - source/
Variation.h , C/C++, 53 lines - source/
bamRemoveDuplicates.cpp , C++, 271 lines - source/
bamRemoveDuplicates.h , C/C++, 10 lines - source/
bamSortByCoordinate.cpp , C++, 97 lines - source/
bamSortByCoordinate.h , C/C++, 11 lines - source/
bam_cat.c , C, 143 lines - source/
bam_cat.h , C/C++, 8 lines - source/
binarySearch2.cpp , C++, 43 lines - source/
binarySearch2.h , C/C++, 7 lines - source/
blocksOverlap.cpp , C++, 41 lines - source/
blocksOverlap.h , C/C++, 10 lines - source/
extendAlign.cpp , C++, 93 lines - source/
extendAlign.h , C/C++, 6 lines - source/
funCompareUintAndSuffixe , C++, 40 liness.cpp - source/
funCompareUintAndSuffixe , C/C++, 11 liness.h - source/
funCompareUintAndSuffixe , C++, 33 linessMemcmp.cpp - source/
funCompareUintAndSuffixe , C/C++, 10 linessMemcmp.h - source/
funPrimaryAlignMark.cpp , C++, 43 lines - source/
funPrimaryAlignMark.h , C/C++, 5 lines - source/
genomeGenerate.h , C/C++, 4 lines - source/
genomeParametersWrite.cp , C++, 45 linesp - source/
genomeParametersWrite.h , C/C++, 9 lines - source/
genomeSAindex.cpp , C++, 217 lines - source/
genomeSAindex.h , C/C++, 11 lines - source/
genomeScanFastaFiles.cpp , C++, 92 lines - source/
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htslib/ , C/C++, 255 lineshtslib/ synced_bcf_reader.h - source/
htslib/ , C/C++, 52 lineshtslib/ tbx.h - source/
htslib/ , C/C++, 824 lineshtslib/ vcf.h - source/
htslib/ , C/C++, 15 lineshtslib/ vcf_sweep.h - source/
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opal/ , C++, 1,568 linesopal.cpp - source/
opal/ , C/C++, 171 linesopal.h - source/
opal/ , C/C++, 5,590 linessimde_avx2.h - source/
outputSJ.cpp , C++, 163 lines - source/
outputSJ.h , C/C++, 4 lines - source/
readBarcodeLoad.h , C/C++, 9 lines - source/
readLoad.cpp , C++, 100 lines - source/
readLoad.h , C/C++, 12 lines - source/
samHeaders.cpp , C++, 108 lines - source/
samHeaders.h , C/C++, 10 lines - source/
serviceFuns.cpp , C++, 351 lines - source/
signalFromBAM.cpp , C++, 209 lines - source/
signalFromBAM.h , C/C++, 13 lines - source/
sjAlignSplit.cpp , C++, 15 lines - source/
sjAlignSplit.h , C/C++, 9 lines - source/
sjdbBuildIndex.cpp , C++, 333 lines - source/
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sjdbInsertJunctions.cpp , C++, 102 lines - source/
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sjdbLoadFromFiles.cpp , C++, 27 lines - source/
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soloInputFeatureUMI.cpp , C++, 44 lines - source/
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twoPassRunPass1.h , C/C++, 11 lines - LICENSE, License, 21 lines
- README.md, Text, 115 lines
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
What the map holds:
- 1 repository of the authors' code, each at its verified commit, with its license and how the link was found in the paper;
- 329 scripts, each with its path and the digest of its content;
- no match between paragraphs and code yet;
- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
No dataset and no data link were found in the paper.
Data and code availability
The dataset generated during this study is available at GEO: GSE307577.
Reproduced under the paper's license (CC BY), from the paper cited above.
Versions
The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.
Version 1, 30 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 5 authors, 4 keywords, 7 MeSH terms, 2 funders, 16 references, 10 RRIDs.
Cite
This paper
Engelenburg, H. J., Smolders, J., Huitinga, I., Hamann, J., & Hsiao, C.-C. (2026). Protocol for isolating viable human central nervous system T cells. STAR protocols, 7(2), 104464. https://
BibTeX
@article{engelenburg2026
author = {Engelenburg, Hendrik J. and Smolders, Joost and Huitinga, Inge and Hamann, Jörg and Hsiao, Cheng-Chih},
title = {{Protocol for isolating viable human central nervous system T cells}},
journal = {STAR protocols},
year = {2026},
month = mar,
volume = {7},
number = {2},
pages = {104464},
publisher = {Elsevier},
issn = {2666-1667},
doi = {10.1016/
url = {https://
pmid = {41886460},
pmcid = {PMC13052091}
}
RIS
TY - JOUR
AU - Engelenburg, Hendrik J.
AU - Smolders, Joost
AU - Huitinga, Inge
AU - Hamann, Jörg
AU - Hsiao, Cheng-Chih
TI - Protocol for isolating viable human central nervous system T cells
T2 - STAR protocols
J2 - STAR Protoc
PY - 2026
DA - 2026/
VL - 7
IS - 2
SP - 104464
SN - 2666-1667
PB - Elsevier
DO - 10.1016/
UR - https://
LA - en
ER -
CSL-JSON
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}
The tracing map gets a citation of its own once an author has validated it and it has a DOI.
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