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| Status |
Public on Mar 01, 2020 |
| Title |
EBF1-mutant bone marrow stroma confers long-term changes in hematopoietic stem cell potential |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Crosstalk between mesenchymal stromal cells (MSCs) and hematopoietic stem and cells (HSCs) is essential for hematopoietic homeostasis and lineage output. Ebf1-deficient MSCs have reduced mesenchymal lineage potential. Ebf1 deletion in Cxcl12-abundant reticular (CAR) cells and PDGFRα+Sca1+CD45-CD31-Lin- (PαS) cells in the bone marrow decreased the numbers of HSPCs and myeloid cells. EBF1 in the bone marrow niche regulates a transcriptional program that determines the functional interactions with HSCs and the long-term balance between the myeloid and lymphoid cell fates.
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| Overall design |
Intact bones (femora and tibia) were chopped into small fragments and crushed in FACS buffer (PBS-2% FCS) using pestle and mortar. For the MSC cells isolation, the bone chips were digested with 0.3% collagenase type II and 0.02% DNase I at 37°C for 45 min with gentle shaking. After enzymatic digestion, the cells were filtered, washed with FACS buffer, and stained accordingly. CAR cells were identified as CD45-CD31-Lin-PDGFRα+Sca1-, PαS cells were identified as CD45-CD31-Lin-PDGFRα+ Sca1+. BD FACSAria Fusion was used for fluorescence-activated cell sorting.
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| Contributor(s) |
Derecka M, Ramamoorthy S, Grosschedl R |
| Citation(s) |
32066955 |
| |
| Submission date |
Mar 06, 2019 |
| Last update date |
Mar 30, 2020 |
| Contact name |
Senthilkumar Ramamoorthy |
| E-mail(s) |
senthilkumar@ie-freiburg.mpg.de
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| Phone |
00497615108731
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| Organization name |
Max Planck Institute of Immunobiology and Epigenetics
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| Department |
Cellular & Molecular Immunology
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| Lab |
Laboratory Rudolf Grosschedl
|
| Street address |
Stübeweg 51
|
| City |
Freiburg |
| ZIP/Postal code |
D-79108 |
| Country |
Germany |
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| Platforms (2) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
| GPL21493 |
Illumina HiSeq 3000 (Mus musculus) |
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| Samples (20)
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| GSM3659297 |
CAR cells - Ebf1+/+,Prx1+/cre – wt Replicate 1 |
| GSM3659298 |
CAR cells - Ebf1+/+,Prx1+/cre – wt Replicate 2 |
| GSM3659299 |
CAR cells - Ebf1+/+,Prx1+/cre – wt Replicate 3 |
| GSM3659300 |
CAR cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 1 |
| GSM3659301 |
CAR cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 2 |
| GSM3659302 |
CAR cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 3 |
| GSM3659303 |
PaS cells - Ebf1+/+,Prx1+/cre – wt Replicate 1 |
| GSM3659304 |
PaS cells - Ebf1+/+,Prx1+/cre – wt Replicate 2 |
| GSM3659305 |
PaS cells - Ebf1+/+,Prx1+/cre – wt Replicate 3 |
| GSM3659306 |
PaS cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 1 |
| GSM3659307 |
PaS cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 2 |
| GSM3659308 |
PaS cells – Ebf1fl/fl;Prx1+/cre – ko Replicate 3 |
| GSM3659309 |
CAR cells – Ebf1+/+ Replicate 1 |
| GSM3659310 |
CAR cells – Ebf1+/+ Replicate 2 |
| GSM3659311 |
CAR cells – Ebf1-/- Replicate 1 |
| GSM3659312 |
CAR cells – Ebf1-/- Replicate 2 |
| GSM3659313 |
PaS cells – Ebf1+/+ Replicate 1 |
| GSM3659314 |
PaS cells – Ebf1+/+ Replicate 2 |
| GSM3659315 |
PaS cells – Ebf1-/- Replicate 1 |
| GSM3659316 |
PaS cells – Ebf1-/- Replicate 2 |
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| Relations |
| BioProject |
PRJNA525888 |
| SRA |
SRP187777 |
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