| RESEARCH PAPER
|Year : 1998 | Volume
| Issue : 3 | Page : 195-198
Cultivation of mouse bone marrow-derived mast cells using Concanavalin-A stimulated splenocyte supernatant
Singh Rashmi, PP Gupta, PP Singh
Objective: To develop mast cells in vitro from mouse bone marrow using concanavalin-A stimulated splenocyte supernatant.
Methods: Bone marrow cells (BMCs) were cultured in 1:1 (vol/vol) mixture of enriched medium (RPMI-1640 with 10% fetal calf serum with L-glutamine, 2 mM HEPES buffer, NaHCO3 (2.0 gm/L) and gentamicin (40 mg/L) and conditioned medium (conmed) derived from concanavalin-A (Con-A; 2.5 'g/ml) stimulated mouse splenocyte.
Results: After culturing mouse bone marrow cells for seven days approximately 1 O-l 5% of cells began to show metachromatic granules with toluidine blue which suggested that these were mast cells. After 14 days of culture around 74-80% cells were identified as mast cells and heterogenous with respect to density of metachromatic granules. Maximum differentiation of bone marrow cells into mast cells were observed when conditioned medium and fresh medium dilution was 1:1 (vol/vol). when dilution of conditioned medium was 16x, 32x. 64x and 128x, lesser conversion of BMCs into mast cells were observed.
Conclusion:The development of mast cells from bone marrow precursor cells in the presence of Con-A stimulated splenocyte supernatant suggested that T-cell derived growth factor had differentiated bone marrow cells into mast cells in vitro when its optimal concentration was used.
Source of Support: None, Conflict of Interest: None