Pre-osteoblast MC3T3-E1 cells (from the cell bank of the Chinese Academy of Sciences [Shanghai, China]) were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA). Osteoblastic differentiation of MC3T3-E1 cells was induced by supplementing a mixture of 10% FBS, dexamethasone (10−8 mol/L), L-ascorbic acid (10 mM), and β-glycerophosphate (10 mM) (Takara Bio, Shiga, Japan) every 2 days.
β glycerophosphate
Manufactured by Takara Bio
Sourced in Japan
β-glycerophosphate is a common organic compound used in various laboratory applications. It serves as a source of phosphate ions, which are essential for numerous biochemical reactions and processes. This compound is widely utilized in cell culture media and as a buffering agent to maintain appropriate pH levels in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dexamethasone (dex), by Takara Bio (3 mentions)
L ascorbic acid, by Takara Bio (3 mentions)
Ascorbic acid, by Takara Bio (3 mentions)
Hydrocortisone, by Takara Bio (3 mentions)
Dulbecco modified eagle medium (dmem), by Takara Bio (2 mentions)
Gentamicin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (2 mentions)
Fetal bovine serum (fbs), by Takara Bio (1 mentions)
Honokiol, by MedChemExpress (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Mc3t3 e1, by Merck Group (1 mentions)
Osteoblast inducer reagent, by Takara Bio (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Mc3t3 e1 subclone 14, by American Type Culture Collection (1 mentions)
Mc3t3 e1 cell, by American Type Culture Collection (1 mentions)
6 protocols using β glycerophosphate
1
MC3T3-E1 Osteoblastic Differentiation
2
Honokiol Promotes Osteoblastic Differentiation
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The pre-osteoblast MC3T3-E1 was obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10 % fetal bovine serum (FBS; Gibco, CA, USA). Osteoblastic differentiation of MC3T3-E1 cells was induced by adding a mixture of 10 % FBS, L-ascorbic acid (10 mM), dexamethasone (10− 8 mol/L) and β-glycerophosphate (10 mM) (Takara Bio, Shiga, Japan) every 2 days. Meanwhile, honokiol was obtained from MedChemExpress (MCE; 10 nM, Shanghai, China). In addition, it was dissolved in 10 mL DMSO and diluted with purified water (1:100).
3
Osteogenic Differentiation of MG63 Cells
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Human preosteoblast MG63 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Tokyo, Japan) with 10% (v/v) fetal bovine serum (FBS; HyClone, Logan, UT, USA) and 1% (v/v) penicillin/streptomycin (PS; Invitrogen). All cultures were maintained at 37°C in a humidified incubator and with 5% CO2. Upon reaching confluence, the cells were cultured in DMEM with FBS supplemented with osteogenic induction medium (OIM) containing ascorbic acid, dexamethasone and β-glycerophosphate (TaKaRa, Tokyo, Japan). The cells were incubated in the OIM-containing culture medium with or without the salmon DNA fragment (100 μg/mL) for 0–21 days.
4
Culturing and Differentiating MC3T3-E1 Osteoblasts
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The clonal mouse osteoblastic cell line, MC3T3-E1 subclone 14, was purchased from American Type Culture Collection (Manassas, VA, USA). The MC3T3-E1 cells were cultured at 37°C in 5% CO2 in Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) with 10% fetal bovine serum (Hyclone; GE Healthcare Life Science, Logan, UT, USA), 100 IU/ml penicillin and 100 µg/ml streptomycin. To induce osteoblast differentiation, MC3T3-E1 cells were cultured at 37°C in 5% CO2 in DMEM with osteoblast inducer reagent containing 1% L-ascorbic acid, 2% β-glycerophosphate and 0.2% hydrocortisone (Takara Bio, Inc., Otsu, Japan). The medium was changed every other day.
5
Osteogenic Differentiation of MC3T3-E1 Cells
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MC3T3-E1 cells (ATCC, Manassas, VA, United States) were used as pre-osteoblasts. MC3T3-E1 cells were cultured for 3 days in culture dishes containing DMEM (Wako) supplemented with 10% FBS (Invitrogen) and antibiotics (0.2 mg/ml gentamicin) (Gibco). Osteogenic differentiation was investigated by supplementing media with 50 mg/L ascorbic acid, 10 μM hydrocortisone, and 10 mM β-glycerophosphate (Takara Bio, Shiga, Japan) (Aaron et al., 2010 (link)).
6
Osteoblast Differentiation Using BMP-2
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MC3T3-E1 cells (1.0 × 105 cells/well) were cultured onto 48-well plates, and treated with DMEM (Wako) supplemented with 10% FBS (Invitrogen), antibiotics (0.2 mg/ml gentamicin) (Gibco), 50 mg/L ascorbic acid, hydrocortisone, and 10 mM β-glycerophosphate (Takara Bio, Shiga, Japan) in the presence and absence of TER (0.1–10 μM) and recombinant mouse bone morphogenetic protein (BMP)-2 (50 ng/ml) (BioLegend, San Diego, CA, United States) for 21 days. Prior to staining, all cells were washed with PBS and fixed with 4% paraformaldehyde (PFA). Then, the cells were stained with an Alizarin red staining kit (Cosmo Bio Co., Ltd., Tokyo, Japan) and an ALP Staining Kit (Fuji Films Co., Tokyo, Japan), following the manufacturer’s instructions (Aaron et al., 2010 (link)).
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