An established murine osteocyte cell line (MLO-Y4) (Kerafast, Boston, MA) was maintained as a subconfluent monolayer culture in the MEM alpha medium (Gibco, Tokyo, Japan) supplemented with 10% fetal calf serum 10 (link). Mouse MSC derived from the bone marrow (Cyagen, Silicon Valley, CA) was maintained in the stem cell growth medium (Cyagen). Both cell lines were cultured at 37°C under 20% O2 and 5% CO2. At the culture reached 80% confluency while being cultured at 37°C under 20% O2 and 5% CO2, both cell lines, MLO-Y4 and MSC, were treated under three different conditions for 24 hours: exposed to Dexamethasone (Dex, MSD, Tokyo, Japan) at the concentration of 0.4 ng/ml (Dex group); hypoxia at a 1% oxygen concentration (Hypoxia group) or both (Dex/Hypoxia group). As a control (C group), both cell lines were cultured under 20% oxygen in the culture medium without Dexamethasone. After the exposure, the numbers of viable and nonviable cells determined using 0.25% trypan blue dye exclusion method were counted with Countess 2 FL (Thermo Fisher Scientific, Waltham, MA), and survival rates were calculated.
Stem cell growth medium
Manufactured by Cyagen
Sourced in Canada, United States
Stem cell growth medium is a specialized cell culture medium designed to support the growth and maintenance of stem cells. It provides the necessary nutrients and growth factors required for stem cells to proliferate and retain their undifferentiated state.
Automatically generated - may contain errors
2 protocols using stem cell growth medium
1
Effects of Dexamethasone and Hypoxia on Osteocyte and Mesenchymal Stem Cell Viability
2
Osteogenic Differentiation of hMSCs with HA-Au Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hMSCs were purchased from Cyagen Biosciences, USA. The hMSCs were plated in 75 cm2 tissue culture flasks and cultured in the stem cell growth medium (Cyagen Biosciences, USA) at 37°C with 5% CO2. The medium was replaced every 3 days during cell culturing. After growing to sub-confluence, cells were collected using Trypsin-EDTA (0.25%), centrifuged, and resuspended in growth medium for reseeding in new culture flasks. Cells were used for further study after two to five passages. For osteogenic induction, cells were trypsinized and cultured in the growth medium overnight. After cells adherence, the medium was changed to an osteogenic induction medium (Cyagen Biosciences, USA) containing phosphate buffer saline (PBS), HA or HA-Au nanoparticles, respectively. For the mechanism experiments, the hMSCs were pretreated with 10 μM ICG-001, one of the Wnt/β-catenin signaling pathway inhibitors, and subsequently cultured with HA-Au nanoparticles in osteoinductive medium.
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!