P2 BMSCs with good growth conditions were cultured in 12-well plates at a density of 5 × 105. After the BMSCs reached 80% confluence, they were incubated from basic medium to chondrogenic differentiation medium [high-glucose DMEM (Corning, USA) with the addition of 10% FBS, 1% P/S, 40 mg/mL proline (Solarbio, Beijing, China), 100 mg/mL sodium pyruvate (Solarbio, China), 50 mg/mL vitamin C (Sigma Aldrich, St. Louis, MO, USA), 50 mg/mL insulin–transferrin–selenium (ThermoFisher Scientific, Waltham, MA, USA), and 10 ng/mL transforming growth factor (Abbkine, Wuhan, China)] [16 (link)]. The chondrogenic induction medium was changed every 2 days for 7 and 14 days.
Proline
Manufactured by Solarbio
Sourced in China
Proline is a high-performance lab equipment designed for protein analysis. It offers accurate and reliable results for protein quantification and purification.
Automatically generated - may contain errors
Lab products found in correlation
Sodium pyruvate, by Solarbio (2 mentions)
Cystine, by Solarbio (2 mentions)
Isoleucine, by Solarbio (2 mentions)
Methionine, by Solarbio (2 mentions)
Glutamine, by Solarbio (2 mentions)
Glycine, by Solarbio (2 mentions)
Tyrosine, by Solarbio (2 mentions)
Histidine, by Solarbio (2 mentions)
Phenylalanine, by Solarbio (2 mentions)
Serine, by Solarbio (2 mentions)
Asparagine, by Solarbio (2 mentions)
Glutamic acid, by Solarbio (2 mentions)
Leucine, by Solarbio (2 mentions)
Valine, by Solarbio (2 mentions)
Transferrin, by Solarbio (2 mentions)
Insulin transferrin selenium, by Thermo Fisher Scientific (1 mentions)
High glucose dmem, by Corning (1 mentions)
Vitamin c, by Merck Group (1 mentions)
2 chloro l phenylalanine, by Adamas (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
5 protocols using proline
1
Chondrogenic Differentiation of BMSCs
2
Ziziphus jujuba Leaf Metabolite Profiling
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Leaves from the Ziziphus jujuba Mill. cv. Dongzao were used, and the picking standard is two leaves and one bud on the fruit-bearing shoot (Figure 1C
3
Chondrogenic Differentiation of MSCs
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The chondrogenic differentiation of MSCs is stimulated by incubation of cells in a medium including ascorbate (Sigma, USA), dexamethasone (Sigma, USA), TGF‐β3 (Yeasen, China), sodium pyruvate (Solarbio, China), selenous acid (Solarbio, China), transferrin (Solarbio, China), proline (Solarbio, China), and L‐glutamine (Solarbio, China). The medium was replaced every third day until day 21 and then the pellets were harvested for further experiments. The glycosaminoglycan content was measured by alcian blue staining. The slides were fixed with 4% PFA and stained with alcian blue for 15 min. Finally, the slides were detected by a microscope (Zeiss, Germany).
4
Bioactive Compound Analytical Methods
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1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Ferric-tripyridyltriazine (Fe3+-TPTZ) is purchased from Beyotime Biotechnology (Shanghai, China). Chromatographic grade analytical standards asparaginic acid, glutamic acid, asparagine, serine, glutamine, histidine, glycine, threonine, citrulline, arginine, alanine, tyrosine, cystine, valine, methionine, tryptophan, phenylalanine, isoleucine, leucine, lysine, hydroxyproline, proline, 5′-CMP, 5′-UMP, 5′-GMP, 5′-IMP, 5′-XMP, 5′-AMP, trehalose, mannitol, quininic acid, l -malic acid and fumaric acid purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). α-Amylase, α-glucosidase, 4-nitrophenyl α-d-glucopyranoside (PNPG), and soluble starch were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). All other reagents were analytical grade and obtained from Chengdu Kelong Chemical Factory (Chengdu, China).
5
Enzymatic Activity Assays Optimization
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β-Galactosidase (β-Gal), horseradish peroxidase, alkaline phosphatase, papain, pepsin, and α-mannosidase were purchased from Beyotime (Shanghai, China). Lysozyme, glucose, bovine serum albumin, γ-globulin, transferrin, mannitol tryptophan and proline were purchased from Solarbio (Beijing, China). Ciprofloxacin, ampicillin, kanamycin, dimethyl sulfoxide (DMSO), Polypropylene pyrrolidone K30 (PVP-K30, MW = 40,000) and all other chemicals and solvents were purchased from Sigma-Aldrich (St. Louis, United States). Luria-Bertani broth (LB) medium was purchased from BD Biosciences (San Jose, CA). All aqueous solutions were prepared with Milli-Q water (≥18 MΩ, Milli-Q, Millipore).
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