Cryo-SFI Serum-Free Freezing Medium -100ml

Cryo-SFI Serum-Free Freezing Medium is a completely ready-to-use reagent: serum-free, animal-derived proteins free, effectively avoiding virus and mycoplasma contamination for your valued cell lines and primary cells specimen. Levels of viability and percent attachment (adherent cells) that are comparable to cells preserved in DMSO and FBS. Serum-Free Cell Freezing Medium can be used for both cells cultured in serum-supplemented growth medium as well as cells grown under serum-free conditions.

1. Serum-free, animal-derived protein-free, effectively avoiding virus and mycoplasma contamination.
2. Easy to use, no need for gradient cooling down for cryopreservation.
3. Compatible with both -80 degree refrigerator and liquid nitrogen cryopreservation.
4. Compatible with well plates (such as 96-well plates, and 48-well plates) and cryovials for cryopreservation.

Product Quality

Long cryopreservation period and high cell viability

Precautions and Disclaimer

This medium is intended for Research & Development use only. It can NOT be used for any animal or human therapeutic treatment, any animal or human consumption, or any diagnostic use.

This medium has been used in the freezing of human 293 HEK cells and CHO stable cell lines.

1. Large-scale rapid micro-cryopreservation of candidate clones of hybridoma cell lines in 96-well plates
2. Serum-free cryopreservation of 293 HEK cells and CHO stable cell lines
3. Routine cryopreservation of passaged cell lines

Protocol

When cell cultures are actively growing for cryopreservation

Conventional cell cryopreservation method [Cryopreservation tube method]

Selecting cryopreserved cells in the exponential growth phase helps to increase the viability of recovered cells.

1. Collect suspension cells or adherent cells according to common methods, place them in centrifuge tubes, and count the cells.
2. Collect the cultured cells by centrifugation (reference centrifugation conditions: 1,000~2,000 rpm, RT, 3~5 min)
3. Add an appropriate amount of serum-free cell freezing solution to the centrifuge tube (cell concentration 5×10⁵ to 5×10⁶ cells/ml). Mix gently.
4. Aliquot the cell mixture into fully labelled cryopreservation tubes.
5. Directly put the cryo-vial containing the cell mixture into the -80°C freezer for long-term cryopreservation.
6. For liquid nitrogen storage, please completely frozen cryo-vials (at least 24h in the -80°C freezer), then tranfer cryo-vials -196°C liquid nitrogen tank.

Cryopreserved cell recovery method [Cryopreservation vial]

1. 1. Remove the cell cryopreservation vial from the freezer or liquid nitrogen tank. Immediately place it in a 37°C water bath at for rapid thawing.
2. After the cell mixture is completely thawed, immediately add 1 ml of cell culture medium to the cells, then transfer the cell mixture into a centrifuge tube containing 4 times the volume of cell culture medium and mix well.
3. Collect the cultured cells by centrifugation (1,000~2,000 rpm, RT, 3~5 min), and completely remove the centrifugation supernatant.
4. Add the appropriate amount of fresh cell culture medium, Incubate cells at 37℃，5%CO₂ for further culture and use.

In situ cryopreservation method [culture plate cryopreservation]

For adherent cells:

1. When the cells in the well plate grow well and the confluence reaches more than 50%, carefully aspirate the medium supernatant from the cell culture plate.
2. Add an appropriate amount of serum-free cell cryopreservation solution into the culture plate to fully infiltrate the cells (the volume is determined by the well plate, generally 100 ul for 96-well plate, 200 ul for 48-well plate).
3. Seal plate with films (Genfne Cat#. ) to prevent any contamination.
4. Put the plate directly into the -80℃ freezer for long-term storage.

For suspension cells:

1. When the cells in the well plate grow well and the density reaches 5X10⁵ to 5X10⁶ cells/ml, centrifuge with a horizontal centrifuge (1,000~2,000 rpm, RT, 3~5 min), carefully aspirate the supernatant from the plate.
2. Add an appropriate amount of the serum-free cell cryopreservation solution into the culture plate to the cells (generally 100 ul for 96-well plate, 200 ul for the 48-well plate).
3. Seal plate with films (Genfne Cat#. ) to prevent any contamination.
4. Put the plate directly into the -80℃ freezer for long-term storage.

In situ cryopreservation recovery method [well plate cryopreservation]

For adherent cells:

1. Remove the frozen cell culture plate from the freezer and thaw it immediately in a 37°C incubator.
2. After the cell cryopreservation solution is completely thawed, carefully aspirate the liquid and add an appropriate amount of cell culture medium immediately.
3. Incubate cells at 37℃，5%CO₂ for further culture and use.

Adherent cells：

1. Take cells from frozen storage and immediately thaw in a 37℃ incubator
2. Centrifugate at 1,000~2,000 rpm, RT, 3~5 min. Carefully remove the supernatant and gently add complete growth medium and mix the pellet cells
3. Incubate cells at 37℃，5%CO₂ for further culture and use.

Storage Conditions

Sorted at 2-8℃

Attention: Freezing- thaw cycling may cause deterioration of the products.

Reference:

1. A novel serum-free method for culturing human prenatal retinal pigment epithelial cells

Invest Ophthalmol Vis Sci . 2008 Feb;49(2):788-99. doi: 10.1167/iovs.07-0777.

2. Cryopreservation of cells using defined serum-free cryoprotective agents. The journal Current Directions in Biomedical Engineering. https://doi.org/10.1515/cdbme-2016-0070

Downloadable Instruction or Test Report

1. Cryo-SFI Serum-free cell cryopreservation 100ml_Instructions-Genfine
2. Cryo-SFI serum-free cell cryopreservation solution Flyer-Genfine