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Comments:
The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a （C57BL/6J X C3H/HeJ）F1 mouse. The cells express laminin and collagen IV, but do not express collagen I or Factor VIII. The cells will support human and murine myelopoiesis in long term culture, and will support certain murine stromal cell dependent pre-B cell lines in vitro.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C Atmosphere: air, 95%; carbon dioxide （CO2）, 5%
Subculturing:
Protocol: Remove medium, and rinse with 0.25% trypsin, 0.53 mM EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature （or at 37C） until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subc*tion Ratio: A subc*tion ratio of 1:6 to 1:10 is recommended Medium Renewal: Every 2 to 3 days
Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2001 recommended serum:ATCC 30-2020
References:
22620: Sutherland HJ, et al. Differential regulation of primitive human hematopoietic cells in long- term cultures maintained on genetically engineered murine stromal cells. Blood 78: 666-672, 1991. PubMed: 1713512 22769: Lemoine FM, et al. Partial characterization of a novel stromal cell-derived pre-B-cell growth factor active on normal and immortalized pre-B cells. Exp. Hematol. 16: 718-726, 1988. PubMed: 3261251 23070: Lemoine FM, et al. Autocrine production of pre-B-cell stimulating activity by a variety of transformed murine pre-B-cell lines. Cancer Res. 48: 6438-6443, 1988. PubMed: 3263188 23353: Lemoine FM, et al. Transformation-associated alterations in interactions between pre-B cells and fibronectin. Blood 76: 2311-2320, 1990. PubMed: 2147863
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