Cell culture medium for differentiation of mesenchymal cells into osteogenic lineages.
Unit100 ml
Cat.No.
C-28013
229,00 €
Currently In stock
We offer a complete Mesenchymal Stem Cell Media System including growth media, differentiation media and human mesenchymal stem cells (MSC).
We offer five MSC Differentiation Media to efficiently induce differentiation of MSC into adipogenic, chondrogenic (with and without inducers), osteogenic or neurogenic lineages, respectively.
Our Mesenchymal Stem Cell Osteogenic Differentiation Media was developed for the directed differentiation of Human Mesenchymal Stem Cells (MSC) from bone marrow, the umbilical cord matrix (Wharton's Jelly) and adipose tissue into osteogenic lineages.
Although all our media are optimized for use with primary human cells, we have received feedback from customers that this particular medium can also be used for porcine cells.
Figure 1.hMSC-BM after in vitro differentiation into osteoblasts using our Mesenchymal Stem Cell Osteogenic Differentiation Medium (Alizarin Red S staining).
The differentiation rate into the osteogenic lineage is 70-100%.
Yes the specialized PromoCell media already contain the optimal amount of L-glutamine. Please don't add extra L-glutamine as this can be toxic for the cells.
FCS is a natural product consisting of many different components like salts hormones vitamins trace elements proteins and enzymes. There can be large lot-to-lot variations between different serum batches regarding the concentrations of growth-promoting factors. The higher the serum content in a culture medium the higher the impact of the variations on the cell culture system. For this reason PromoCell has developed several serum-reduced and serum-free media where part of the serum has been replaced by more defined factors like cytokines hormones or vitamins.Media with reduced serum concentrations have the benefit that they produce more standardized culture conditions over a long time span.
Serum turbidity is usually caused by cryoprecipitation of lipid components during freezing and thawing. The more times the serum is subjected to freeze/thaw cycles the more turbidity is noticed. It can be minimized by freezing the serum in aliquots at -20°C and thawing these aliquots individually at the time of use.
A concentration > 10% FBS is needed to completely inactivate the trypsin. As most of PromoCell's growth media are serum-reduced or serum-free; the use of a trypsin inhibitor like TNS is highly recommended.
The practice of heat inactivation was originally developed when only serum from adult animals was available. Adult serum contains high serum complement which may destroy cells under certain conditions. Heating serum (30 min 56°C) is intended to inactivate the complement. Today serum is often heat-inactivated without any evidence of beneficial effect. When using FCS (fetal calf serum) heat inactivation is not necessary for most cell lines or cell types. PromoCell does not use heat-inactivated FCS for the preparation of their growth media.
If antibiotics are deemed necessary the recommended final concentrations are:
100 U/ml penicillin + 100 µg/ml streptomycin or
50 µg/ml gentamicin + 50 ng/ml amphotericin B
Please note: Addition of antibiotics can reduce the growth rate of the cells.
After addition of the SupplementMix or SupplementPack to our basal medium do I have to add FCS you obtain the complete growth medium. No further supplementation with serum or growth factors is required. Please note: Our media do not contain antibiotics. If you wish to use antibiotics you can add penicillin/streptomycin or gentamicin/amphotericin B at standard concentrations. Addition of antibiotics can however reduce the doubling time of the cells up to 30-40%.
Yes you can aliquot the SupplementMix upon delivery and freeze down 2 or 4 individual aliquots at -20°C. This way you can prepare smaller volumes (2 x 250 ml or 4 x 125 ml) of complete culture medium and thus extend the time you can use the medium.
Upon arrival the basal media should be stored between 4°C and 8°C the supplements at -20°C.
Please do not freeze our cell culture media. Freezing can lead to irreversible precipitation of media components and the quality can no longer be assured.
The qualitative and quantitative composition of the supplements can be found on our website and in the data sheets of the specialized media. When there is no such information specified; the composition of the supplements is confidential.
The formulation of our basal media is proprietary information. If you need to know the concentration of a particular component for your experiments please contact the PromoCell Technical Customer Service.
It has been shown that phenol red has estrogenic properties. Phenol red-free media are therefore generally used in studies evaluating steroid hormone action in cultured estrogen-responsive cells (Berthois et al. 1986). Furthermore phenol red can also interfere with some analytical methods like photometric analyses.
The Growth Medium Kit allows customization of the end concentrations of growth supplements. It is therefore more flexible than the Medium "ready-to-use" and can eg. be used to prepare a starvation medium.
Please note: Modification of supplement concentrations may have an impact on cell growth. You should test in advance whether and for how long the cells can survive the altered culture conditions.
Human Mesenchymal Stem Cells have retained the ability to differentiate into a variety of cell types including fat cells; chondrocytes; and osteoblasts. PromoCell supply hMSC from 3 different tissues: bone marrow (hMSC-BM; C-12974); umbilical cord matrix (hMSC-UC; C-12971); and adipose tissue (hMSC-AT; C-12977).
MSC-BM show very good differentiation into bone cells but also into chondrocytes and fat cells when the respective Differentiation Media is used.
MSC-UC have a high potential to differentiate into chondrocytes but only weak potential for fat or bone cell differentiation.
In contrast; MSC-AT differentiate very well into fat and bone cells but only moderately into chondrocytes.
In other words; to obtain a high percentage of bone cells; MSC-BM or MSC-AT are the cells of choice. There are of course lot-to-lot variations and the differentiation will decrease from passage to passage. If you need MSCs with a particularly high osteoblast differentiation capacity; you can call our Technical Customer Service before placing your order so that we can select an appropriate cell lot for you.
Adipogenic differentiation can be identified morphologically and without any staining by the formation of intracellular lipid vesicles.
In contrast; when MSC differentiate into bone cells; there is no significant change in morphology. It is recommended to perform Alkaline Phosphatase staining to detect osteoblastic differentiation or Alizarin Red S staining to show osteoblast mineralization.
Chondrogenic differentiation is generally performed as spheroids in 3-D cell culture and not in 2-D monolayer culture. Staining with Alcian Blue to visualize the differentiation process is indispensable.
Neurogenic differentiation can be detected using neuron specific markers (e.g. beta-3 tubulin; NeuN; MAP2) and by their typical neuronal morphology.
For differentiation protocols; please see attached Application Notes.
We have tested the differentiation capacity of our hMSC into adipocytes chondrocytes and osteoblasts over time and still see good differentiation rates after 10 population doublings i.e. at passage 5. However the differentiation potential declines with ongoing population doublings. To obtain optimal differentiation rates experiments should be performed as early in culture as possible.
PromoCell Cell Culture Media "ready-to-use" consist of basal medium and SupplementMix.
PromoCell Culture Media Kits consist of basal medium and SupplementPack.
Addition of the supplements (SupplementMix or SupplementPack; respectively) to the appropriate basal medium will result in identical growth media.
Osteogenic differentiation of hMSCs with PromoCell MSC Osteogenic Differentiation Medium takes ∼12-14 days (please view the respective Application Note for a detailed protocol). Use fibronectin-coated plates and change the medium every third day.
The bone cells tend to detach from the plastic after approximately 2 weeks; when differentiation is complete. Therefore; the tests should be performed promptly and the cells should be maintained in MSC Osteogenic Differentiation Medium until then.
The PBS buffering enhances the Alizarin Red staining (precipitation of the dye) and makes it more intense.
Leave the PBS on the cells after staining/washing and analyze the sample immediately; as the dye may bleed upon prolonged storage without embedding.
Metabolic Activity During Mesenchymal Stem cell (MSC) Differentiation
Mesenchymal stem cells: why optimizing manufacturing processes is key for a successful application
Tools for mesenchymal stem cell culture
MSC reproducibility: Towards the standardization of Mesenchymal Stem Cells
Osteogenic differentiation and analysis of mesenchymal stem cells (MSC)
Cells in Action: Mitosis in Human Mesenchymal Stem Cells (MSCs)
Automated monitoring of metabolic activity and differentiation of human mesenchymal stem cells
Real-time analysis of stem cell proliferation during differentiation
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