2021-01-25
ENA first public
2021-01-15
ENA last update
ERC000011
ENA-CHECKLIST
SAMEA7819882
External Id
Dept. of Hematology Erasmus MC
INSDC center alias
Dept. of Hematology Erasmus MC
INSDC center name
2021-01-25T04:05:29Z
INSDC first public
2021-01-15T09:21:22Z
INSDC last update
public
INSDC status
ERS5567085
SRA accession
E-MTAB-10020:19-MI17170-02M_MSC_TOMpos
Submitter Id
3
week
age
ArrayExpress
http://www.ebi.ac.uk/efo/EFO_0009737
broker name
bone marrow stromal cell
cell type
house mouse
common name
Protocols: 3 or 11 week-old VE-Cadherin-Cre;LoxP-tdTomato mice were euthanized by cervical dislocation. Collagenased bone fraction was obtained from long bones and hips. Cells were stained with the following antibodies: anti-CD45.2 (clone 104, 1:100), anti-Ter119 (clone TER-119, 1:25), anti-CD144 (clone BV13, 1:200), anti-CD51 (clone RMV-7, 1:50), anti-Sca1 (clone D7, 1:50) all from Biolegend, anti-CD140α (clone APA5, 1:100) from eBioscience, and anti-CD31 (clone MEC 13.3, 1:100) from BD Biosciences. Alexa Fluor™ 488-conjugated Streptavidin (1:100, Invitrogen) was used to detect cells marked by biotin anti-CD51 antibody. 7-Amino-Actinomycin D viability dye (7-AAD, 1:100, Beckman Coulter) was used to distinguish between living and dead cells. BMSCs (CD45-TER119-CD31-CD144-SCA-1+ CD51+) and OLCs (CD45-TER119-CD31-CD144-Sca1-CD51+) were directly sorted in Trizol. RNA of sorted cells was extracted using TRIzol according to the manufacturer’s instructions for RNA, in combination isolation with GenElute LPA (Sigma-Aldrich). cDNA was prepared using the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech) for Illumina Sequencing.
description
adult
http://www.ebi.ac.uk/efo/EFO_0001272
developmental stage
VE-Cadherin-Cre-positive;LoxP-tdTomato-positive
genotype
CD45-TER119-CD31-CD144-SCA-1+ CD51+
immunophenotype
3W02-MSC-POS
individual
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
Mus musculus
http://purl.obolibrary.org/obo/NCBITaxon_10090
organism
bone
http://purl.obolibrary.org/obo/UBERON_0002481
organism part
male
http://purl.obolibrary.org/obo/PATO_0000384
sex
B6;129-Tg(Cdh5-cre)1Spe/J; Rosa26-tdTomato (B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J)
strain
19-MI17170-02M_MSC_TOMpos
title