M2-10B4

The M2-10B4 cell line is a clone derived from bone marrow stromal cells from a (C57BL/6J X C3H/HeJ)F1 mouse. These stromal cells are essential components of the bone marrow microenvironment and play a significant role in supporting hematopoiesis. The M2-10B4 cells are particularly valuable for research focused on the interactions between stromal and hematopoietic cells, as they can support both human and murine myelopoiesis in long-term culture. Additionally, these cells can sustain certain murine stromal cell-dependent pre-B cell lines in vitro, making them a versatile tool in hematopoietic research.

The M2-10B4 cells express important extracellular matrix components such as laminin and collagen IV, which contribute to their ability to support hematopoietic cells. However, they do not express collagen I or Factor VIII, which distinguishes them from other stromal cell lines. The presence of laminin and collagen IV is critical for the maintenance of the bone marrow microenvironment, influencing cell adhesion, differentiation, and signaling pathways. Researchers often utilize the M2-10B4 cell line in co-culture systems to explore the effects of stromal cells on the behavior of hematopoietic progenitors, particularly in the context of bone marrow physiology and disease models.

Given their origin and functional properties, M2-10B4 cells are an essential model for studying the bone marrow niche, especially in relation to hematological disorders such as leukemia. They are also useful in drug screening and the development of therapeutic strategies targeting the bone marrow microenvironment.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.