Na citrate

Manufactured by AppliChem

Na-citrate is a chemical compound used in various laboratory applications. It functions as a buffer solution, maintaining a specific pH level in aqueous environments. The core purpose of Na-citrate is to provide a controlled chemical environment for experiments and analyses.

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Lab products found in correlation

Alcian blue, by Merck Group (4 mentions) Papain, by Merck Group (4 mentions) Chondroitin sulphate, by Merck Group (4 mentions) Ascorbic acid 2 phosphate, by Merck Group (3 mentions) Dexamethasone (dex), by Merck Group (3 mentions) Insulin transferrin selenium x, by Thermo Fisher Scientific (3 mentions) Sodium chloride (nacl), by AppliChem (2 mentions) Dmem glutamax, by Thermo Fisher Scientific (2 mentions) Ethylene diamine tetraacetic acid (edta), by AppliChem (2 mentions) Tgf β1, by Thermo Fisher Scientific (2 mentions) L proline, by Merck Group (1 mentions) Transforming growth factor β1, by Thermo Fisher Scientific (1 mentions) Sybr green, by Thermo Fisher Scientific (1 mentions) Calf thymus dna standard, by Merck Group (1 mentions)

4 protocols using na citrate

Chondrogenic Differentiation of MSCs in Collagen Scaffolds

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Collagen type I cubes (Biopad, Euroresearch, Italy) were used as a scaffold to support cellular differentiation [31 (link)]. MSC (8 × 10⁵) were seeded per cube and kept for 30 min to allow adhesion before the addition of chondrogenic medium. MSC-collagen constructs were cultured for 3 weeks in chondrogenic medium consisting of advanced DMEM + GlutaMAX (Gibco), 2.5% FBS, 100 units/mL penicillin, 100 mg/mL streptomycin, 2.5 μg/mL amphotericin B, 40 ng/mL dexamethasone (Sigma), 50 μg/mL ascorbic acid 2-phosphate (Sigma), 35 μg/mL l-proline (Sigma), 1× Insulin-Transferrin-Selenium X (Gibco), and 10 ng/ml transforming growth factor (TGF)-β1 (Peprotech).
Glycosaminoglycan (GAG) accumulation was determined as a chondrogenic marker. GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of two constructs per sample at 60 °C with 125 μg/mL papain (Sigma-Aldrich) in 5 mM l-cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl, and 5 mM EDTA (all AppliChem). GAG accumulation was determined by binding to alcian blue (Fluka, Sigma) and quantified using chondroitin sulphate (Sigma) reference standards [32 (link)].

Bertolo A., Capossela S., Fränkl G., Baur M., Pötzel T, & Stoyanov J. (2017). Oxidative status predicts quality in human mesenchymal stem cells. Stem Cell Research & Therapy, 8, 3.

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Quantifying GAG Accumulation and DNA Levels

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GAG accumulation was quantified with alcian blue binding assay after 6 h digestion of three constructs per sample at 60°C with 125 µg/ml papain (Sigma) in 5 mM L-cysteine-HCl (Fluka-Sigma), 5 mM Na-citrate, 150 mM NaCl, 5 mM EDTA (all AppliChem). GAG accumulation was determined by binding to alcian blue (Sigma), absorption was measured at 595 nm and quantified against chondroitin sulphate (Sigma) reference standards^{18 (link)}. Total double stranded DNA was measured for each sample after papain digestion. The amount of DNA was determined using SYBR green (Invitrogen) fluorescent assay (absorption measured at 535 nm), quantified by referring to calf thymus DNA standards (Sigma).

Steffen F., Bertolo A., Affentranger R., Ferguson S.J, & Stoyanov J. (2018). Treatment of Naturally Degenerated Canine Lumbosacral Intervertebral Discs with Autologous Mesenchymal Stromal Cells and Collagen Microcarriers: A Prospective Clinical Study. Cell Transplantation, 28(2), 201-211.

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Chondrogenic Differentiation of MSCs on Collagen Scaffolds

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Collagen type I cubes (Biopad, Euroresearch, Italy) were used as scaffold to support cellular differentiation⁵⁶. MSC (5 × 10⁵ cells) were seeded onto collagen cubes and kept for 30 minutes to allow adhesion, before addition of chondrogenic medium. MSC-collagen constructs were cultured for three weeks at 5% O₂ in chondrogenic medium, consisting of: advanced DMEM + GlutaMAX (Gibco), 2.5% FBS, 100 units/mL penicillin, 100 mg/mL streptomycin, 2.5 µg/mL amphotericin B, 40 ng/mL dexamethasone (Sigma), 50 µg/mL ascorbic acid 2-phosphate (Sigma), 1x Insulin-Transferrin-Selenium X (Gibco), and 10 ng/ml transforming growth factor-β1 (TGF-β1, Peprotech).
Glycosaminoglycan (GAG) accumulation was determined as chondrogenic marker. GAG accumulation was quantified with alcian blue binding assay after six hours digestion of three cell-constructs per sample at 60 °C with 125 µg/ml papain (Sigma) in 5 mM L-cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl and 5 mM EDTA (all AppliChem). GAG accumulation was determined by binding to alcian blue (Fluka, Sigma) and quantified using chondroitin sulphate (Sigma) reference standards^{57 (link)}.

Bertolo A., Baur M., Guerrero J., Pötzel T, & Stoyanov J. (2019). Autofluorescence is a Reliable in vitro Marker of Cellular Senescence in Human Mesenchymal Stromal Cells. Scientific Reports, 9, 2074.

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Chondrogenic Differentiation of MSCs on Collagen Scaffolds

Cited 1 time

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Collagen type I cubes (Biopad, Euroresearch, Italy) were used as scaffold to support cellular differentiation^{17 (link),46}. MSC (5 × 10⁵ cells) were seeded onto collagen cubes and kept for 30 min to allow adhesion, before addition of chondrogenic medium. MSC-collagen constructs were cultured for three weeks at 5% O₂ in chondrogenic medium, consisting of: advanced DMEM + GlutaMAX (Gibco), 2.5% FBS, 100 units/mL penicillin, 100 mg/mL streptomycin, 2.5 µg/mL amphotericin B, 40 ng/mL dexamethasone (Sigma), 50 µg/mL ascorbic acid 2-phosphate (Sigma), 1 × Insulin-Transferrin-Selenium X (Gibco), and 10 ng/ml transforming growth factor-β1 (TGF-β1, Peprotech).
Glycosaminoglycan (GAG) accumulation was determined as chondrogenic marker. GAG accumulation was quantified with alcian blue binding assay after six hours digestion of three cell-constructs per sample at 60 °C with 125 µg/ml papain (Sigma) in 5 mM L-cysteine-HCl (Fluka), 5 mM Na-citrate, 150 mM NaCl and 5 mM EDTA (all AppliChem). GAG accumulation was determined by binding to alcian blue (Fluka, Sigma) and quantified using chondroitin sulphate (Sigma) reference standards^{47 (link)}.

Bertolo A., Guerrero J, & Stoyanov J. (2020). Autofluorescence-based sorting removes senescent cells from mesenchymal stromal cell cultures. Scientific Reports, 10, 19084.

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