EB3 Cells

3D cell culture, Immunology

The EB3 cell line is a mouse embryonic stem (ES) cell line that has been extensively studied for its response to cell–cell contact. One of the key features of the EB3 line is its expression of connexin30.3 (Cx30.3), a connexin isoform specifically expressed in the undifferentiated state of these cells. Cx30.3 expression is highly responsive to cell–cell contact, as demonstrated in experiments where hanging drop culture, which increases cell–cell interaction, led to a significant 1.73-fold increase in Cx30.3 expression compared to single cell cultures.
This contact-dependent upregulation of Cx30.3 is mediated through the E-cadherin signaling pathway, and knockdown experiments with ß-catenin, a downstream factor of cadherin signaling, have confirmed its involvement. Inhibition of E-cadherin or β-catenin led to a reduction in Cx30.3 expression, indicating that cell–cell contact activation of Cx30.3 is at least partially regulated by this pathway. Interestingly, this cell contact response is specific to connexin30.3, as the expression of connexin43 (Cx43), another connexin isoform, did not exhibit the same level of sensitivity to cell contact in these experiments.
The EB3 cell line is a useful model for studying the mechanisms of cell–cell communication and the role of gap junctions in early embryonic development. The unique expression pattern of Cx30.3 in response to contact suggests that it may play a role as a signaling mediator during tissue morphogenesis and differentiation in the undifferentiated stem cell state.

Suspension

Burkitt lymphoma

Male

Lymphoblast

HLA A3, Aw32, Cw2

RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)

Gently homogenize the cell suspension in the flask by pipetting up and down, then take a representative sample to determine the cell density per ml. Dilute the suspension to achieve a cell concentration of 1 x 10^5 cells/ml with fresh culture medium, and aliquot the adjusted suspension into new flasks for further cultivation.

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

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.

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.

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

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.

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

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.

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

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

Bone