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Freezing Medium Cryo-SFM
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商品描述
Freezing Medium Cryo-SFM，C-29910，PromoCell：
產(chǎn)品介紹：
PromoCell Cryo-SFM is a protein-free, defined and animal component-free cryopreservation medium. The optimized formulation is based on methylcellulose, DMSO and other cryoprotectants for the cryopreservation of human and animal primary cells. Cryo-SFM provides excellent results with all types of cells including primary human cells, stem cells and cell lines. Cells frozen in Cryo-SFM exhibit superior viability, attachment and subsequent growth performance after thawing. The medium is the first choice for all types of primary cells and cell lines.
常見問(wèn)題：
1.What’s the difference between PromoCell’s Cryo-SFM and a standard freezing medium?
Most standard freezing media contain 20%-90% FCS and 10% DMSO. PromoCell Cryo-SFM does not contain any serum and is free of animal components. It contains DMSO, methylcellulose and other cryoprotective components instead of FCS or BSA. Since many PromoCell culture media have a low serum content or are serum-free, our Cryo-SFM is ideally suited to cryopreserve these cultures using serum-free conditions.
2.What is the advantage of Cryo-SFM compared to the standard freezing media?
In a serum-reduced or serum-free cell culture system, it is essential to cryopreserve the cells in a serum-free environment. When using a serum-containing freezing medium, the serum exerts long lasting effects on the culture system. This influence can only be removed by dilution through many subsequent passages. The use of Cryo-SFM will therefore lead to much more standardized culture conditions as it does not contain serum.
3.What is the DMSO concentration in Cryo-SFM?
The DMSO concentration in our Cryo-SFM is 7.5%.
4.How often should I change the growth medium when culturing PromoCell Normal Human Cells?
Generally, the medium should be changed every 2-3 days. Please note: Following thawing, the first medium change should be performed after 16-24 hours to prevent cell damage due to residual freezing medium.
5.Can I re-freeze PromoCell Normal Human Cells?
It is possible to re-freeze PromoCell Normal Human Cells. But you should keep in mind that each additional freezing cycle leads to a loss in proliferation potential and cell number. This means that the number of population doublings (PDs) specified in the CoA can no longer been guaranteed. If you still want to re-freeze the cells, please do it at a low passage number and use PromoCell Cryo-SFM (C-29910), a serum-free freezing medium which we also use in our cell culture lab.
6.What is the difference between cryopreserved and the proliferating cells?
Our cryopreserved cells are frozen in Cryo-SFM (C-29910) and are shipped on dry ice. After delivery, they must be transferred into liquid nitrogen or can be defrosted directly into an appropriate tissue culture vessel using the recommended Growth Media.
Proliferating cells are defrosted at PromoCell and shipped as growing cultures in T25 flasks (T75 for Mesenchymal Stem Cells and Pericytes). Upon arrival, they usually have a density between 60-90%.
7.Is it feasible to store the melanocytes for a second time in liquid nitrogen after subculture, similar to cell lines?
Upon arrival, you can either defrost the cells immediately and seed them in a TC vessel or store the vial in liquid nitrogen until needed.
It is possible to re-freeze normal human cells but PromoCell does not recommend it, because each freezing cycle leads to a loss in proliferation potential. If you want to freeze them, please use Cryo-SFM (C-29910) which is a serum-free freezing medium that prevents the possibility that any serum is introduced into your serum-free cell culture system. It is also recommended to freeze the cells at an early passage.
8.Can I use the PromoCell growth medium to prepare my own freezing medium?
No, we do not recommend to freeze our growth media as they are very complex in composition. Their freezing can lead to irreversible precipitations. We either recommend to use standard DMEM [complete freezing medium: 70% DMEM, 20% FCS, 10% DMSO] or our serum-free Cryo-SFM (# C-29910). When using a low serum or serum-free cell culture system, cryopreservation in a serum-free freezing medium like Cryo-SFM is highly recommended.
9.Could you please let me know whether you have used the baculovirus expression system to produce components of your media?
Baculovirus is generally used in conjunction with insect cells (Sf-9, Sf-21) to produce recombinant proteins (cytokines, growth factors).
None of our Specialized Media (Media for Primary Human Cells; Blood and Stem Cell Media) contain recombinant proteins produced in insect cells. This also applies to our Cryo-SFM Freezing Medium.
10.What are the key points relating to proliferation, differentiation and culturing of HWP?
PromoCell’s Normal Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations. The cells are frozen in our serum-free freezing medium (Cryo-SFM) at the end of passage 1 (= secondary culture). Two randomly selected vials are then subjected to quality control, which includes e.g. determination of growth characteristics, control of morphology, and tests for differentiation capacity into mature adipocytes.
The recommended seeding density of preadipocytes after thawing/trypsinization is 5,000 cells/cm2; cells should be trypsinized before reaching 90% confluence. Population doubling times are usually between 20-50 hrs (10 population doublings guaranteed). Using a 1:4 split ratio, you can perform 4-5 passages with the cells.
Preadipocyte Growth Medium (C-27417) is used to propagate the cells. To induce differentiation of preadipocytes into adipocytes, cells are grown in PromoCell’s Preadipocyte Growth Medium until they reach 100% confluency. Cells are then cultured in Preadipocyte Differentiation Medium (C-27437) for 72 h, followed by 10-14 days in Adipocyte Nutrition Medium (C-27439). During this time the cells start to accumulate droplets of fat which can be visualized under the microscope.
We recommend performing differentiation experiments at population doubling numbers lower than 4-5, in order to reach a high differentiation level of the culture.
PromoCell Cryo-SFM is a protein-free, defined and animal component-free cryopreservation medium. The optimized formulation is based on methylcellulose, DMSO and other cryoprotectants for the cryopreservation of human and animal primary cells. Cryo-SFM provides excellent results with all types of cells including primary human cells, stem cells and cell lines. Cells frozen in Cryo-SFM exhibit superior viability, attachment and subsequent growth performance after thawing. The medium is the first choice for all types of primary cells and cell lines.
常見問(wèn)題：
1.What’s the difference between PromoCell’s Cryo-SFM and a standard freezing medium?
Most standard freezing media contain 20%-90% FCS and 10% DMSO. PromoCell Cryo-SFM does not contain any serum and is free of animal components. It contains DMSO, methylcellulose and other cryoprotective components instead of FCS or BSA. Since many PromoCell culture media have a low serum content or are serum-free, our Cryo-SFM is ideally suited to cryopreserve these cultures using serum-free conditions.
2.What is the advantage of Cryo-SFM compared to the standard freezing media?
In a serum-reduced or serum-free cell culture system, it is essential to cryopreserve the cells in a serum-free environment. When using a serum-containing freezing medium, the serum exerts long lasting effects on the culture system. This influence can only be removed by dilution through many subsequent passages. The use of Cryo-SFM will therefore lead to much more standardized culture conditions as it does not contain serum.
3.What is the DMSO concentration in Cryo-SFM?
The DMSO concentration in our Cryo-SFM is 7.5%.
4.How often should I change the growth medium when culturing PromoCell Normal Human Cells?
Generally, the medium should be changed every 2-3 days. Please note: Following thawing, the first medium change should be performed after 16-24 hours to prevent cell damage due to residual freezing medium.
5.Can I re-freeze PromoCell Normal Human Cells?
It is possible to re-freeze PromoCell Normal Human Cells. But you should keep in mind that each additional freezing cycle leads to a loss in proliferation potential and cell number. This means that the number of population doublings (PDs) specified in the CoA can no longer been guaranteed. If you still want to re-freeze the cells, please do it at a low passage number and use PromoCell Cryo-SFM (C-29910), a serum-free freezing medium which we also use in our cell culture lab.
6.What is the difference between cryopreserved and the proliferating cells?
Our cryopreserved cells are frozen in Cryo-SFM (C-29910) and are shipped on dry ice. After delivery, they must be transferred into liquid nitrogen or can be defrosted directly into an appropriate tissue culture vessel using the recommended Growth Media.
Proliferating cells are defrosted at PromoCell and shipped as growing cultures in T25 flasks (T75 for Mesenchymal Stem Cells and Pericytes). Upon arrival, they usually have a density between 60-90%.
7.Is it feasible to store the melanocytes for a second time in liquid nitrogen after subculture, similar to cell lines?
Upon arrival, you can either defrost the cells immediately and seed them in a TC vessel or store the vial in liquid nitrogen until needed.
It is possible to re-freeze normal human cells but PromoCell does not recommend it, because each freezing cycle leads to a loss in proliferation potential. If you want to freeze them, please use Cryo-SFM (C-29910) which is a serum-free freezing medium that prevents the possibility that any serum is introduced into your serum-free cell culture system. It is also recommended to freeze the cells at an early passage.
8.Can I use the PromoCell growth medium to prepare my own freezing medium?
No, we do not recommend to freeze our growth media as they are very complex in composition. Their freezing can lead to irreversible precipitations. We either recommend to use standard DMEM [complete freezing medium: 70% DMEM, 20% FCS, 10% DMSO] or our serum-free Cryo-SFM (# C-29910). When using a low serum or serum-free cell culture system, cryopreservation in a serum-free freezing medium like Cryo-SFM is highly recommended.
9.Could you please let me know whether you have used the baculovirus expression system to produce components of your media?
Baculovirus is generally used in conjunction with insect cells (Sf-9, Sf-21) to produce recombinant proteins (cytokines, growth factors).
None of our Specialized Media (Media for Primary Human Cells; Blood and Stem Cell Media) contain recombinant proteins produced in insect cells. This also applies to our Cryo-SFM Freezing Medium.
10.What are the key points relating to proliferation, differentiation and culturing of HWP?
PromoCell’s Normal Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations. The cells are frozen in our serum-free freezing medium (Cryo-SFM) at the end of passage 1 (= secondary culture). Two randomly selected vials are then subjected to quality control, which includes e.g. determination of growth characteristics, control of morphology, and tests for differentiation capacity into mature adipocytes.
The recommended seeding density of preadipocytes after thawing/trypsinization is 5,000 cells/cm2; cells should be trypsinized before reaching 90% confluence. Population doubling times are usually between 20-50 hrs (10 population doublings guaranteed). Using a 1:4 split ratio, you can perform 4-5 passages with the cells.
Preadipocyte Growth Medium (C-27417) is used to propagate the cells. To induce differentiation of preadipocytes into adipocytes, cells are grown in PromoCell’s Preadipocyte Growth Medium until they reach 100% confluency. Cells are then cultured in Preadipocyte Differentiation Medium (C-27437) for 72 h, followed by 10-14 days in Adipocyte Nutrition Medium (C-27439). During this time the cells start to accumulate droplets of fat which can be visualized under the microscope.
We recommend performing differentiation experiments at population doubling numbers lower than 4-5, in order to reach a high differentiation level of the culture.
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