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The Cytokine Mix M1 and M2 should not be subjected to further freeze/thaw cycles.
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Based on negative feedback we have received from customers using bead baths, we strongly discourage the use of bead baths to thaw our cells. It can lead to reduced viability or significantly slower growing cells. If you don’t have a “normal” water bath but only a bead bath in your lab, thaw the vial in a beaker of water in the bead bath. Ensure the water is heated to exactly 37°C using a thermometer placed in the warmed water. Be sure to hold the vial in your hand, and not in a floater, as described in the thawing protocol.
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To avoid the spreading of the matrix and to have nice drop-like domes, it is crucial to preheat the plate for 2h at 37°C and to work fast when transferring the gel to the wells. Placing a warming plate under your laminar air flow hood when transferring the gel-cell-mix to the preheated plate may help. From our testing, we found that Nunclon Sphera wells were not ideal for our organoid system as the dome did not stay adhered, but it was able to move on the bottom and the dome became misshaped overtime. However, the altered shape of the dome did not affect the organoids and were successfully cultured for 4 weeks. If movement is not an issue for the user, we recommend that extra caution be taken during media changes by holding the plate at an angle and carefully pipette to remove old medium. Aspirating with vacuum suction can damage or destroy the dome.
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Human mesenchymal stem cells (MSCs) are able to attach without fibronectin coating when cultured in MSC Adipogenic Differentiation Medium 2.Learn more
However, under these conditions the risk of cell detachment during adipogenic differentiation is significantly higher.
Recommendation
To ensure stable cell attachment throughout the differentiation process, we strongly recommend using fibronectin‑ or vitronectin‑coated tissue culture vessels.
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PromoCell growth media are developed and optimized for the in vitro-culture of human primary cells. However, many of our media have also been successfully used for:Learn more
- Primary cells from other species
- Different primary cell types
- Immortalized cell lines
Please note: Culture performance may vary depending on the cell type and application. We therefore recommend reviewing the available publications and adapting the culture conditions as needed for your specific cell type and experimental setup.
Our Scientific Support team will be happy to assist you in identifying relevant publications and evaluating their relevance for your particular cell type and application.