Differentiating intermediate mesoderm cells from 4 independent differentiation experiments were cryopreserved on day 7. Cells were dissociated after CHIR pulse using Trypsin–EDTA, and single cells were counted and resuspended in ice-cold Nutrifreez (Biological Industries). Cryovials containing 8 × 106 cells/mL were rate controlled (− 1 °C/min) frozen to − 80 °C. 24 h later vials were transferred and stored in liquid nitrogen. For thawing, vials were warmed at 37 °C and cells were transferred to a 50 mL tube containing plain DMEM and counted. Cells were centrifuged at the previously indicated density, resuspended in a small volume of 10% FBS in DMEM and pipetted onto the transwell membrane to allow tissue sheet formation using the templates. The protocol was continued as described above until day 7 + 18.
Nutrifreez
Manufactured by Sartorius
Nutrifreez is a laboratory equipment designed for the efficient freezing and storage of biological samples. It provides controlled temperature environments to preserve the integrity of materials, ensuring reliable results for various research and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nutrifreez
1
Cryopreservation and Thawing of Mesoderm Cells
2
Cryopreservation and Tissue Engineering
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Differentiating intermediate mesoderm cells from 4 independent differentiation experiments were cryopreserved on day 7. Cells were dissociated after CHIR pulse using Trypsin-EDTA, and single cells were counted and resuspended in ice-cold Nutrifreez (Biological Industries). Cryovials containing 8 x 10 6 cells/mL were rate controlled (-1°C/min) frozen to -80°C. 24 h later vials were transferred and stored in liquid nitrogen. For thawing, vials were warmed at 37°C and cells were transferred to a 50 mL tube containing plain DMEM and counted. Cells were centrifuged at the indicated density, resuspended in a small volume of 10% FBS in DMEM and pipetted onto the transwell membrane to allow tissue sheet formation using the templates. The protocol was continued as described above until day 7+18.
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