Macrophage Microvesicles Induce Macrophage Differentiation and miR-223 Transfer
We characterized macrophage-derived microvesicles and whether they influenced the differentiation of naïve monocytes. We also identified the miRNA content of the macrophage-derived microvesicles.
Microvesicles bind to and activate target cells.
All experiments were done in triplicates. Primary monocytes were collected from buffy coats (BC). The freshly isolated monocytes from three donors (Mono1-3) were either treated with GM-CSF or subjected to RNA isolation. Following treatment, MVs were isolated from the GM-CSF-treated macrophage cultures. RNA was isolated from the remaining cells for profiling (GM1-3). The isolated MVs were then used to treat new BC monocytes for 24 h (BC-GMCSF-MV24). A fraction of the new BC monocytes was subjected to RNA extraction for profiling (BC1-3). For THP1 cells, they were treated with either DMSO or PMA to produce MVs. The MVs were collected and the remaining cells lyzed for RNA extraction and profiling (DMSO1-3 and PMA1-3). The collected MVs from the DMSO or PMA-treated THP1 cells were incubated with new THP1 for 24 h and designated DMSO-MV24 or PMA-MV24. We had a total of 24 samples.
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We used GeneChip microarrays to examine changes in gene expression induced by MV in primary monocyte-derived macrophages (MDM) and in THP1 cells, and compare this to cells treated with GM-CSF and PMA, respectively.
| Sample ID | !Sample_title | treatment | cell type | cell group | Conditions |
|---|---|---|---|---|---|
| GSM1026969 | Freshly isolated monocytes used as MV donors, repl. 1 | primary monocytes | Monocytes | primary | Monocyte-Mvdon |
| GSM1026970 | Freshly isolated monocytes used as MV donors, repl. 2 | primary monocytes | Monocytes | primary | Monocyte-Mvdon |
| GSM1026971 | Freshly isolated monocytes used as MV donors, repl. 3 | primary monocytes | Monocytes | primary | Monocyte-Mvdon |
| GSM1026972 | GM-CSF-treated macrophages used as MV donors, repl. 1 | differentiated MDM | Monocyte-derived macrophages | primary | MDM + MV |
| GSM1026973 | GM-CSF-treated macrophages used as MV donors, repl. 2 | differentiated MDM | Monocyte-derived macrophages | primary | MDM + MV |
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| Sample ID | !Sample Title | Treatment | Cell type | Cell group | Conditions |
|---|---|---|---|---|---|
| GSM1026969 | Freshly isolated monocytes used as MV donors, repl. 1 |
primary monocytes |
Monocytes |
primary |
Monocyte-Mvdon |
| GSM1026970 | Freshly isolated monocytes used as MV donors, repl. 2 |
primary monocytes |
Monocytes |
primary |
Monocyte-Mvdon |
| GSM1026971 | Freshly isolated monocytes used as MV donors, repl. 3 |
primary monocytes |
Monocytes |
primary |
Monocyte-Mvdon |
| GSM1026972 | GM-CSF-treated macrophages used as MV donors, repl. 1 |
differentiated MDM |
Monocyte-derived macrophages |
primary |
MDM + MV |
| GSM1026973 | GM-CSF-treated macrophages used as MV donors, repl. 2 |
differentiated MDM |
Monocyte-derived macrophages |
primary |
MDM + MV |
| GSM1026974 | GM-CSF-treated macrophages used as MV donors, repl. 3 |
differentiated MDM |
Monocyte-derived macrophages |
primary |
MDM + MV |
| GSM1026975 | Freshly isolated monocytes used as MV recipients, repl. 1 |
differentiated MDM |
Monocytes |
primary |
Monocyte-Mvrep |
| GSM1026976 | Freshly isolated monocytes used as MV recipients, repl. 2 |
differentiated MDM |
Monocytes |
primary |
Monocyte-Mvrep |
| GSM1026977 | Freshly isolated monocytes used as MV recipients, repl. 3 |
differentiated MDM |
Monocytes |
primary |
Monocyte-Mvrep |
| GSM1026978 | Freshly isolated monocyte incubated 24 h with MV from other GM-CSF-treated MDM, repl. 1 |
MDM after interaction with MDM-derived MV |
Monocyte-derived macrophages |
primary |
MDM + GMCSF + MV |
| GSM1026979 | Freshly isolated monocyte incubated 24 h with MV from other GM-CSF-treated MDM, repl. 2 |
MDM after interaction with MDM-derived MV |
Monocyte-derived macrophages |
primary |
MDM + GMCSF + MV |
| GSM1026980 | Freshly isolated monocyte incubated 24 h with MV from other GM-CSF-treated MDM, repl. 3 |
MDM after interaction with MDM-derived MV |
Monocyte-derived macrophages |
primary |
MDM + GMCSF + MV |
| GSM1026981 | THP1+DMSO used as MV donors, repl. 1 |
THP1 cells treated with DMSO |
THP1 cells |
cell line |
THP-1 + DMSO |
| GSM1026982 | THP1+DMSO used as MV donors, repl. 2 |
THP1 cells treated with DMSO |
THP1 cells |
cell line |
THP-1 + DMSO |
| GSM1026983 | THP1+DMSO used as MV donors, repl. 3 |
THP1 cells treated with DMSO |
THP1 cells |
cell line |
THP-1 + DMSO |
| GSM1026984 | THP1 incubated 24 h with MV from THP1-DMSO donors, repl. 1 |
THP1 cells treated with THP1+DMSO-derived MV |
THP1 cells |
cell line |
THP1 +DMSO +MV |
| GSM1026985 | THP1 incubated 24 h with MV from THP1-DMSO donors, repl. 2 |
THP1 cells treated with THP1+DMSO-derived MV |
THP1 cells |
cell line |
THP1 +DMSO +MV |
| GSM1026986 | THP1 incubated 24 h with MV from THP1-DMSO donors, repl. 3 |
THP1 cells treated with THP1+DMSO-derived MV |
THP1 cells |
cell line |
THP1 +DMSO +MV |
| GSM1026987 | THP1+PMA used as MV donors, repl. 1 |
THP1 cells treated with PMA |
THP1 cells |
cell line |
THP-1 + PMA |
| GSM1026988 | THP1+PMA used as MV donors, repl. 2 |
THP1 cells treated with PMA |
THP1 cells |
cell line |
THP-1 + PMA |
| GSM1026989 | THP1+PMA used as MV donors, repl. 3 |
THP1 cells treated with PMA |
THP1 cells |
cell line |
THP-1 + PMA |
| GSM1026990 | THP1 incubated 24 h with MV from THP1-PMA donors, repl. 1 |
THP1 cells treated with THP1+PMA-derived MV |
THP1 cells |
cell line |
THP-1 +PMA +MV |
| GSM1026991 | THP1 incubated 24 h with MV from THP1-PMA donors, repl. 2 |
THP1 cells treated with THP1+PMA-derived MV |
THP1 cells |
cell line |
THP-1 +PMA +MV |
| GSM1026992 | THP1 incubated 24 h with MV from THP1-PMA donors, repl. 3 |
THP1 cells treated with THP1+PMA-derived MV |
THP1 cells |
cell line |
THP-1 +PMA +MV |
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