organism: Mus musculus
description: 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.
attributes.tag: ENA first public
attributes.value: 2021-01-25
attributes.tag: ENA last update
attributes.value: 2021-01-15
attributes.tag: External Id
attributes.value: SAMEA7819886
attributes.tag: INSDC center alias
attributes.value: Dept. of Hematology Erasmus MC
attributes.tag: INSDC center name
attributes.value: Dept. of Hematology Erasmus MC
attributes.tag: INSDC first public
attributes.value: 2021-01-25T04:05:29Z
attributes.tag: INSDC last update
attributes.value: 2021-01-15T09:21:22Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: SRA accession
attributes.value: ERS5567089
attributes.tag: Sample Name
attributes.value: E-MTAB-10020:19-MI17170-03M_MSC_TOMpos
attributes.tag: Submitter Id
attributes.value: E-MTAB-10020:19-MI17170-03M_MSC_TOMpos
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: bone marrow stromal cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: VE-Cadherin-Cre-positive;LoxP-tdTomato-positive
attributes.tag: immunophenotype
attributes.value: CD45-TER119-CD31-CD144-SCA-1+ CD51+
attributes.tag: individual
attributes.value: 3W03-MSC-POS
attributes.tag: organism part
attributes.value: bone
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: B6; 129-Tg(Cdh5-cre)1Spe/J; Rosa26-tdTomato (B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J)
attributes.tag: title
attributes.value: 19-MI17170-03M_MSC_TOMpos