SCM016
Mesenchymal Stem Cell Freezing Medium (1x)
The Mesenchymal Stem Cell Freezing Medium (1x) has been optimized & validated for Stem cell culture. This medium is available in a 50ml format.
Se connecterpour consulter vos tarifs contractuels et ceux de votre entreprise/organisme
About This Item
Code UNSPSC :
12352207
eCl@ss :
32160801
Nomenclature NACRES :
NA.75
Produits recommandés
Niveau de qualité
Forme
liquid
Fabricant/nom de marque
Chemicon®
Technique(s)
cell culture | stem cell: suitable
Entrée
sample type mesenchymal stem cell(s)
Conditions d'expédition
dry ice
Application
1. Thaw cell culture freezing medium completely and mix well by gently swirling bottle. Keep freezing medium on ice during use.
2. Cells to be frozen should be in late log phase growth.
3. Monolayers will need to be dissociated. After dissociation, cells are resuspended in Mesenchymal Stem Cell Expansion Medium (Cat. No. SCM015) and counted to determine viability and number.
4. Centrifuge cells at 300 x g for 3 min. Remove the medium above the pellet.
5. Resuspend the cells in Mesenchymall Stem Cell Freezing Medium at a concentration of ~4 x 106 cells/mL. Freeze 1 mL of cells/vial. After the cells have been resuspended and aliquoted into appropriate cryogenic storage vials, they can be placed in a freezing container and the normal freeze down procedure should be started within five minutes.
6. Cells must be stored at or below -80°C. For long term storage the cells should be stored in ultra-low temperature freezer (-150°C), or in liquid nitrogen (-196°C).
7. Thawing of cryopreserved cells should be as follows:
a. Do not thaw the cells until the recommended medium and appropriate plasticware and/or glassware are on hand.
b. Thaw cells quickly in a 37°C water bath. Important: Do not vortex the cells.
c. Sterilize vial by rinsing with 70% ethanol.
d. In a laminar flow hood, use a 1 or 2 mL pipette to transfer the cells to a sterile 15 mL conical tube. Be careful to not introduce any bubbles during the transfer process.
e. Using a 10 mL pipette, slowly add dropwise 9 mL of Mesenchymal Stem Cell Expansion Medium (pre-warmed at 37°C) to the 15 mL conical tube. IMPORTANT: Do not add the whole volume of medium at once to the cells. This may result in decreased cell viability due to osmotic shock.
f. Gently mix the cell suspension by slow pipeting up and down twice. Be careful to not introduce any bubbles. IMPORTANT: Do not vortex the cells.
g. Centrifuge the tube at 300 x g for 2-3 minutes to pellet the cells.
h. Decant as much of the supernatant as possible
i. Resuspend the cells in a total volume of 10 mL of Mesenchymal Stem Cell Expansion Medium (pre-warmed at 37°C).
j. Plate the cell mixture onto a 10-cm tissue culture plate.
k. Incubate the cells at 37°C in a 5% CO2 humidified incubator.
l. The next day, exchange the medium with fresh Mesenchymal Stem Cell Expansion Medium (pre-warmed to 37°C). Exchange with fresh medium every two to three days thereafter.
m. When the cells are approximately 80% confluent, they can be dissociated with Accutase® (Cat. No. SCR005) and passaged or alternatively frozen for later use.
2. Cells to be frozen should be in late log phase growth.
3. Monolayers will need to be dissociated. After dissociation, cells are resuspended in Mesenchymal Stem Cell Expansion Medium (Cat. No. SCM015) and counted to determine viability and number.
4. Centrifuge cells at 300 x g for 3 min. Remove the medium above the pellet.
5. Resuspend the cells in Mesenchymall Stem Cell Freezing Medium at a concentration of ~4 x 106 cells/mL. Freeze 1 mL of cells/vial. After the cells have been resuspended and aliquoted into appropriate cryogenic storage vials, they can be placed in a freezing container and the normal freeze down procedure should be started within five minutes.
6. Cells must be stored at or below -80°C. For long term storage the cells should be stored in ultra-low temperature freezer (-150°C), or in liquid nitrogen (-196°C).
7. Thawing of cryopreserved cells should be as follows:
a. Do not thaw the cells until the recommended medium and appropriate plasticware and/or glassware are on hand.
b. Thaw cells quickly in a 37°C water bath. Important: Do not vortex the cells.
c. Sterilize vial by rinsing with 70% ethanol.
d. In a laminar flow hood, use a 1 or 2 mL pipette to transfer the cells to a sterile 15 mL conical tube. Be careful to not introduce any bubbles during the transfer process.
e. Using a 10 mL pipette, slowly add dropwise 9 mL of Mesenchymal Stem Cell Expansion Medium (pre-warmed at 37°C) to the 15 mL conical tube. IMPORTANT: Do not add the whole volume of medium at once to the cells. This may result in decreased cell viability due to osmotic shock.
f. Gently mix the cell suspension by slow pipeting up and down twice. Be careful to not introduce any bubbles. IMPORTANT: Do not vortex the cells.
g. Centrifuge the tube at 300 x g for 2-3 minutes to pellet the cells.
h. Decant as much of the supernatant as possible
i. Resuspend the cells in a total volume of 10 mL of Mesenchymal Stem Cell Expansion Medium (pre-warmed at 37°C).
j. Plate the cell mixture onto a 10-cm tissue culture plate.
k. Incubate the cells at 37°C in a 5% CO2 humidified incubator.
l. The next day, exchange the medium with fresh Mesenchymal Stem Cell Expansion Medium (pre-warmed to 37°C). Exchange with fresh medium every two to three days thereafter.
m. When the cells are approximately 80% confluent, they can be dissociated with Accutase® (Cat. No. SCR005) and passaged or alternatively frozen for later use.
Forme physique
Serum-containing formulation. Contains 10% DMSO.
Stockage et stabilité
Store at -20°C.
Informations légales
Accutase is a registered trademark of Innovative Cell Technologies, Inc.
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Clause de non-responsabilité
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Code de la classe de stockage
10 - Combustible liquids
Classe de danger pour l'eau (WGK)
WGK 1
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".
Déjà en possession de ce produit ?
Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
Les clients ont également consulté
Notre équipe de scientifiques dispose d'une expérience dans tous les secteurs de la recherche, notamment en sciences de la vie, science des matériaux, synthèse chimique, chromatographie, analyse et dans de nombreux autres domaines..
Contacter notre Service technique