Table 2.
Phenotype of stromal fractions after culture.
| Cell fractionsa,b | Growthc | Phenotype of 28 day stromad |
||||||
|---|---|---|---|---|---|---|---|---|
| Sca1 | gp38 | CD51 | CD105 | ERTR7 | CD140a | Thy1.2 | ||
| Endothelial cell markers | ||||||||
| CD31+ (n = 1) | - | |||||||
| CD31- (n = 1) | ∗∗ | ++ | + | +++ | - | - | + | + |
| VCAM1+ (n = 1) | - | |||||||
| VCAM1- (n = 1) | ∗ | +++ | +++ | +++ | +++ | - | - | +++ |
| PVRC markers | ||||||||
| CD146+ (n = 4) | - | |||||||
| CD146- (n = 4) | - | |||||||
| MadCAM1+ (n = 3) | - | |||||||
| MAdCAM1- (n = 3) | - | |||||||
| Common stromal markers | ||||||||
| Sca-1+ (n = 3) | ∗ | +++ | ++ | +++ | + | - | - | +++ |
| Sca-1- (n = 3) | ∗ | +++ | ++ | +++ | + | - | - | ++ |
| CD51+ (n = 1) | ∗ | +++ | +++ | +++ | + | - | + | +++ |
| CD51- (n = 1) | ∗ | +++ | +++ | +++ | ++ | - | - | +++ |
| ER-TR7+ (n = 2) | ∗ | +++ | ++ | +++ | - | - | - | +++ |
| ER-TR7- (n = 2) | ∗ | +++ | +++ | ++ | +++ | - | - | - |
aStromal cells were isolated from murine spleen using collagenase treatment and stained with antibodies specific for CD45.2 to gate out hematopoietic cells. They were then stained for known markers of endothelial cells (CD31 and VCAM), perivascular reticular cells (PVRC) (CD146, MadCAM1) or common stromal cell markers (Sca-1, CD51 and ER-TR7) to gate distinct fractions. Where indicated, replicate independent sorting experiments were conducted (n). Sorted subsets were cultured in sDMEM for 28 days.b1–3 × 104 cells were plated, except for the MadCAM1+ subset where ∼3.4 × 103 cells were available for plating.cCell growth: confluent by 10 days (∗∗), confluent by 28 days (∗), no confluence (-).dCells were trypsinised and stained with antibodies to determine phenotype. +++, >80% cells expressing; ++, 50–80%; +, 10–50%; -, <10%.