1 × 106 cells were cultured as pellet culture in StemPro Osteocyte/Chondrocyte basal medium (Life Technologies) supplemented with StemPro Chondrogenesis supplement (Life Technologies) in a humidified incubator at 37°C and 5% CO2. Fresh medium was provided every 2–3 days. After 21 days, medium was aspirated and cell pellets were washed with PBS before fixation in 4% PFA for 60 min. After 3 washings steps with ddH2O, Alcian Blue staining solution was added [0.1 mg/ml Alcian Blue dissolved in 6 parts EtOH and 4 parts of acetic acid (Sigma-Aldrich)] and incubated over night at room temperature under exclusion of light. Cell pellets were embedded in O.C.T compound (TAAB Laboratory Equipment Ltd., Reading, UK) and frozen in super-cooled ethanol at −80°C. Cryostat sections (10 μm) were prepared (Bright OT5000 cryostat), mounted on poly-lysine-coated glass slides (VWR) and visualized using a Nikon TiE microscope (Nikon UK Ltd., Kingston upon Thames, UK).
Stempro osteocyte chondrocyte basal medium
Manufactured by Thermo Fisher Scientific
StemPro Osteocyte/Chondrocyte basal medium is a cell culture medium designed to support the growth and maintenance of osteocytes and chondrocytes, which are specialized cells found in bone and cartilage tissue, respectively. The medium provides the necessary nutrients and growth factors to sustain these cell types in vitro.
Automatically generated - may contain errors
2 protocols using stempro osteocyte chondrocyte basal medium
1
Chondrogenic Differentiation of Mesenchymal Cells
2
Multilineage Differentiation of Amniotic Fluid Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AFSCs were seeded at 4000 cells per cm 2 and grown to 95% confluency.
Growth media was then removed, cells were washed once with PBS and cultured for 21 days with Stem XVivo Adipogenic Base Media (R&D Systems) supplemented with Stem XVivo Adipogenic Supplement (100X) according to the suppliers instructions. Cells were stained for 30 minutes with Oil Red O and washed with ddH2O. Osteogenic Differentiation AFSCs were seeded at 4000 cells per cm 2 and grown to 95% confluency.
Growth media was then removed, cells were washed once with PBS and cultured for 21 days with Stem XVivo Osteogenic Base Media (R&D Systems) supplemented with Stem Pro Osteocyte/Chondrocyte basal medium (Life Technologies) according to the supplier's instructions.
Calcium staining was carried out for 30 minutes using Alizarin Red S and washed with ddH 2 O before capturing images.
Chondrogenic Differentiation For chondrogenic differentiation, 1x10 6 AFSCs were pelleted at 300g.
Growth media was then removed, cells were washed once with PBS and cultured for 21 days with Stem XVivo Adipogenic Base Media (R&D Systems) supplemented with Stem XVivo Adipogenic Supplement (100X) according to the suppliers instructions. Cells were stained for 30 minutes with Oil Red O and washed with ddH2O. Osteogenic Differentiation AFSCs were seeded at 4000 cells per cm 2 and grown to 95% confluency.
Growth media was then removed, cells were washed once with PBS and cultured for 21 days with Stem XVivo Osteogenic Base Media (R&D Systems) supplemented with Stem Pro Osteocyte/Chondrocyte basal medium (Life Technologies) according to the supplier's instructions.
Calcium staining was carried out for 30 minutes using Alizarin Red S and washed with ddH 2 O before capturing images.
Chondrogenic Differentiation For chondrogenic differentiation, 1x10 6 AFSCs were pelleted at 300g.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!