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| Status |
Public on Feb 08, 2024 |
| Title |
Osteomacs promote maintenance of murine hematopoiesis through megakaryocyte-induced upregulation of Embigin and CD166 |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Maintenance of hematopoietic stem cell (HSC) function in the niche is an orchestrated event. Osteomacs (OM), are key cellular components of the niche. Previously, we documented that osteoblasts, OM, and megakaryocytes interact to promote hematopoiesis. Here, we further characterize OM and identify megakaryocyte-induced mediators that augment the role of OM in the niche. Single cell mRNAseq, mass spectrometry, and CyTOF examination of megakaryocyte-stimulated OM suggested that upregulation of CD166 and Embigin on OM augment their hematopoiesis maintenance function. CD166 knockout OM or shRNA-Embigin knockdown OM, confirmed that loss of these molecules significantly reduced OM ability to augment the osteoblast-mediated hematopoietic enhancing activity. Recombinant CD166 and Embigin partially substituted for OM function, characterizing both proteins as critical mediators of OM hematopoietic function. Our data identify Embigin and CD166 as OM-regulated critical components of HSC function in the niche and potential participants in various in vitro manipulations of stem cells.
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| Overall design |
1x105 neonatal calvarial cells (NCC) were cultured for 16hrs with or without 50,000 megakaryocytes were stained with CD45, F4/80, and CD41. Sorted CD45+F4/80+CD41- cells were suspended at 3x105 cells/ml PBS then dispensed into medium sized IFC chips (Fluidigm C1 Single-Cell Auto Prep System)(Zhang et al., 2019). mRNA was isolated followed by cDNA synthesis (Clontech SMART-Se v4 Ultra Low Input RNA Kit). Up to 0.4ng cDNA was used for library preparation and indexing with Nextera XT DNA Library Prep Kit (Illumina, Inc., San Diego, CA). Pooled libraries (4nM, quality checked) were used for 150b paired-end sequencing (NextSeq 500). Cells that passed quality testing were analyzed.
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| Contributor(s) |
Mohamad SF, Srour EF, Zhang C |
| Citation(s) |
38458190 |
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| Submission date |
Feb 03, 2024 |
| Last update date |
May 30, 2024 |
| Contact name |
Safa F Mohamad |
| E-mail(s) |
safa24oct@gmail.com
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| Organization name |
Boston Children's Hospital/ Harvard Medical School
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| Street address |
125 Park Drive, Apt 15
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02215 |
| Country |
USA |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (44)
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| GSM8061880 |
OM (NCC+MK), 20 replicates, scRNA-seq 1 |
| GSM8061881 |
OM (NCC+MK), 20 replicates, scRNA-seq 2 |
| GSM8061882 |
OM (NCC+MK), 20 replicates, scRNA-seq 3 |
| GSM8061883 |
OM (NCC+MK), 20 replicates, scRNA-seq 4 |
| GSM8061884 |
OM (NCC+MK), 20 replicates, scRNA-seq 5 |
| GSM8061885 |
OM (NCC+MK), 20 replicates, scRNA-seq 6 |
| GSM8061886 |
OM (NCC+MK), 20 replicates, scRNA-seq 7 |
| GSM8061887 |
OM (NCC+MK), 20 replicates, scRNA-seq 8 |
| GSM8061888 |
OM (NCC+MK), 20 replicates, scRNA-seq 9 |
| GSM8061889 |
OM (NCC+MK), 20 replicates, scRNA-seq 10 |
| GSM8061890 |
OM (NCC+MK), 20 replicates, scRNA-seq 11 |
| GSM8061891 |
OM (NCC+MK), 20 replicates, scRNA-seq 12 |
| GSM8061892 |
OM (NCC+MK), 20 replicates, scRNA-seq 13 |
| GSM8061893 |
OM (NCC+MK), 20 replicates, scRNA-seq 14 |
| GSM8061894 |
OM (NCC+MK), 20 replicates, scRNA-seq 15 |
| GSM8061895 |
OM (NCC+MK), 20 replicates, scRNA-seq 16 |
| GSM8061896 |
OM (NCC+MK), 20 replicates, scRNA-seq 17 |
| GSM8061897 |
OM (NCC+MK), 20 replicates, scRNA-seq 18 |
| GSM8061898 |
OM (NCC+MK), 20 replicates, scRNA-seq 19 |
| GSM8061899 |
OM (NCC+MK), 20 replicates, scRNA-seq 20 |
| GSM8061900 |
OM (NCC+MK), 24 replicates, scRNA-seq 1 |
| GSM8061901 |
OM (NCC+MK), 24 replicates, scRNA-seq 2 |
| GSM8061902 |
OM (NCC+MK), 24 replicates, scRNA-seq 3 |
| GSM8061903 |
OM (NCC+MK), 24 replicates, scRNA-seq 4 |
| GSM8061904 |
OM (NCC+MK), 24 replicates, scRNA-seq 5 |
| GSM8061905 |
OM (NCC+MK), 24 replicates, scRNA-seq 6 |
| GSM8061906 |
OM (NCC+MK), 24 replicates, scRNA-seq 7 |
| GSM8061907 |
OM (NCC+MK), 24 replicates, scRNA-seq 8 |
| GSM8061908 |
OM (NCC+MK), 24 replicates, scRNA-seq 9 |
| GSM8061909 |
OM (NCC+MK), 24 replicates, scRNA-seq 10 |
| GSM8061910 |
OM (NCC+MK), 24 replicates, scRNA-seq 11 |
| GSM8061911 |
OM (NCC+MK), 24 replicates, scRNA-seq 12 |
| GSM8061912 |
OM (NCC+MK), 24 replicates, scRNA-seq 13 |
| GSM8061913 |
OM (NCC+MK), 24 replicates, scRNA-seq 14 |
| GSM8061914 |
OM (NCC+MK), 24 replicates, scRNA-seq 15 |
| GSM8061915 |
OM (NCC+MK), 24 replicates, scRNA-seq 16 |
| GSM8061916 |
OM (NCC+MK), 24 replicates, scRNA-seq 17 |
| GSM8061917 |
OM (NCC+MK), 24 replicates, scRNA-seq 18 |
| GSM8061918 |
OM (NCC+MK), 24 replicates, scRNA-seq 19 |
| GSM8061919 |
OM (NCC+MK), 24 replicates, scRNA-seq 20 |
| GSM8061920 |
OM (NCC+MK), 24 replicates, scRNA-seq 21 |
| GSM8061921 |
OM (NCC+MK), 24 replicates, scRNA-seq 22 |
| GSM8061922 |
OM (NCC+MK), 24 replicates, scRNA-seq 23 |
| GSM8061923 |
OM (NCC+MK), 24 replicates, scRNA-seq 24 |
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| Relations |
| BioProject |
PRJNA1072995 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE254995_Processed_data.txt.gz |
605.8 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
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