2 phospho l ascorbic acid

Manufactured by Merck Group

Sourced in United States

2-phospho-L-ascorbic acid is a chemical compound used in laboratory settings. It functions as a source of ascorbic acid (vitamin C) and a phosphorus-containing compound. This product is intended for research and experimental purposes only.

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Lab products found in correlation

Dexamethasone (dex), by Merck Group (5 mentions) β glycerophosphate, by Merck Group (5 mentions) Trypsin edta, by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (2 mentions) Epidermal growth factor (egf), by R&D Systems (1 mentions) Dexamethasone (dex), by Thermo Fisher Scientific (1 mentions) Platelet derived growth factor, by R&D Systems (1 mentions) Linoleic acid albumin, by Merck Group (1 mentions) Its liquid media supplement, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Collagenase, by Merck Group (1 mentions) Cetylpyridinium chloride, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline solution, by Thermo Fisher Scientific (1 mentions) Alizarin red, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

14 protocols using 2 phospho l ascorbic acid

Isolation and Characterization of MAPCs

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MAPCs were isolated from a human donor through bone marrow aspiration. Cell isolation was processed according to previously described methods.25 (link) Briefly, MAPCs were cultured in fibronectin-coated plastic tissue culture flasks. Cell cultures were maintained under low oxygen tension in a humidified atmosphere of 5% CO₂. Cells were cultured in a media containing low-glucose (D)MEM (Life Technologies, Grand Island, New York, USA) supplemented with fetal bovine serum (Atlas, Fort Collins, Colorado, USA), ITS liquid media supplement (Sigma), MCDB (Sigma), platelet-derived growth factor (R&D Systems, Minneapolis, Minnesota, USA), epidermal growth factor (R&D Systems), dexamethasone, penicillin/streptomycin (Life Technologies), 2-phospho-L-ascorbic acid and linoleic acid-albumin (Sigma). Cells were passaged every 3–4 days and harvested using trypsin/EDTA (Life Technologies). The cells were positive for CD49c and CD90 and negative for MHC class II and CD45 (all antibodies (Abs) were from BD Biosciences, San Jose, CaliforniaA, USA). Cells were cryopreserved in media and 5% dimethyl sulfoxide. Before administration, the MAPCs were counted with trypan blue exclusion, and the final concentration was adjusted according with the percentage of live cells.

Cardenes N., Aranda-Valderrama P., Carney J.P., Sellares Torres J., Alvarez D., Kocyildirim E., Wolfram Smith J.A., Ting A.E., Lagazzi L., Yu Z., Mason S., Santos E., Lopresti B.J, & Rojas M. (2019). Cell therapy for ARDS: efficacy of endobronchial versus intravenous administration and biodistribution of MAPCs in a large animal model. BMJ Open Respiratory Research, 6(1), e000308.

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Characterization of Osteogenic Potential

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All solvents were purchased from Fisher and used as received. Phosphate-buffered saline (PBS) solution, Dulbecco’s modified Eagle’s medium (DMEM), fungizone antimycotic (Fz), penicillin (Pen), streptomycin (Strep) and trypsin-EDTA were supplied by Gibco Life Technologies. Poly(ε-caprolactone) oligomers, hexamethylene diisocyanate, Span^® 80, dibutyltin dilaurate, collagenase, paraformaldehyde (PFA), 2-phospho-l-ascorbic acid, β-glycerophosphate, DAPI, alizarin red, Triton X100 and cetylpyridinium chloride were purchased from Sigma-Aldrich. Fetal bovine serum (FBS) was obtained from PANBiotech GmbH.
Human dermal fibroblasts were isolated, using standard procedures, from foreskins of 4-year-old children with informed consent of the parents. Fibroblasts were used for the experiments between passages 4 and 6. hMSC were obtained by plastic adhesion from bone marrow samples collected from hematologically healthy patients undergoing routine total hip replacement surgery. All samples were obtained after informed consent from donors from the Hôpital d’Instruction des Armées Percy (Clamart, France).

Rohman G., Changotade S., Frasca S., Ramtani S., Consalus A., Langueh C., Collombet J.M, & Lutomski D. (2019). In vitro and in vivo proves of concept for the use of a chemically cross-linked poly(ester-urethane-urea) scaffold as an easy handling elastomeric biomaterial for bone regeneration. Regenerative Biomaterials, 6(6), 311-323.

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Co-differentiation of Mouse BMSCs

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BMSCs were isolated from mouse long bones as described above, plated at 2 × 10⁶ cells per cm² in αMEM supplemented with 15% of FCS and grown until confluent (approximately 10 days) with twice weekly medium changes. After reaching confluence the medium was replaced with co‐differentiation adipogenic and osteogenic medium, αMEM supplemented with 10% of FCS, 2 mM of β‐glycerophosphate, 50 µg/mL of 2‐Phospho‐L‐ascorbic acid trisodium salt, and 100 nM of dexamethasone (Sigma‐Aldrich), adapted from Ghali et al.³⁰ Culture was continued with 3‐weekly changes of medium for 12 more days, plates fixed, and adipocytes stained with Oil‐red‐O. After staining, the plates were imaged using a Nikon Diaphot inverted microscope fitted with a ToupTeK camera, using a 10× lens, and number of adipocytes per image field counted, counting at least three fields for every mouse cell donor.

Johnson de Sousa Brito F.M., Butcher A., Pisconti A., Poulet B., Prior A., Charlesworth G., Sperinck C., Scotto di Mase M., Liu K., Bou‐Gharios G., Jurgen van 't Hof R, & Daroszewska A. (2021). Syndecan‐3 enhances anabolic bone formation through WNT signaling. The FASEB Journal, 35(4), e21246.

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Osteogenic Differentiation of Mesenchymal Stem Cells

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MSC (Lonza) were expanded in Dulbecco’s modified Eagle (DMEM) medium with low glucose (Gibco), supplemented with 10% v/v fetal bovine serum (FBS) qualified for MSC expansion (Gibco), and 1% v/v penicillin/streptomycin (P/S) (Gibco). Cells were used between passages 6 and 10. For OB differentiation, MSC were plated in 24-well plates at a density of 25 000 cells/well. Cultures were left undisturbed for 3 days at 37 °C in a 5% CO₂ humidified incubator to allow MSC to reach confluency. Osteogenesis was induced by replacing the expansion medium with osteogenic medium (OM), composed of DMEM with low glucose (Gibco), supplemented with 10% v/v FBS, 1% v/v P/S, 100 nmol·L⁻¹ Dexamethasone (Sigma-Aldrich), 10 μmol·L⁻¹ β-glycerophosphate (Sigma), and 50 μmol·L⁻¹ 2-phospho-L-ascorbic acid (Sigma-Aldrich). The OM was replaced every 2–3 days for 21 days. To collect the conditioned medium, OB-lineage cells were incubated in 300 µL of OM and left undisturbed for 3 days at 37 °C in a 5% CO₂ humidified incubator. Afterward the medium was filtrated and stored at −80 °C until further use.

Leitão L., Neto E., Conceição F., Monteiro A., Couto M., Alves C.J., Sousa D.M, & Lamghari M. (2020). Osteoblasts are inherently programmed to repel sensory innervation. Bone Research, 8, 20.

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Osteogenic Differentiation of PDLSCs

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The PDLSCs were cultured in α-MEM, DMEM or EHFM until passage 4. Then, the cells were plated at a density of 4000 cell/cm² for histological and molecular analysis. The next day, the media were replaced with osteogenic differentiation media obtained by adding 0.1 μM dexamethasone (no. D4902; Sigma Aldrich), 50 μg/ml 2-phospho-L-ascorbic acid (no. 49752; Sigma Aldrich), and 10 mM β-glycerophosphate (no. G9422; Sigma Aldrich) to α-MEM, DMEM and EHFM. Osteogenic differentiation was also induced in standardized conditions by culturing PDLSCs in a commercial osteogenic differentiation medium (no. A1007201; StemPro™ Osteogenesis Differentiation Kit, Life Technologies).

Di Vito A., Giudice A., Chiarella E., Malara N., Bennardo F, & Fortunato L. (2018). In Vitro Long-Term Expansion and High Osteogenic Potential of Periodontal Ligament Stem Cells: More Than a Mirage. Cell Transplantation, 28(1), 129-139.

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Synthesis of Nanoparticle Conjugates

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Tetraethylorthosilicate (TEOS), 3-aminopropyltriethoxysilane (APTS), silver nitrate (AgNO₃), Tetrakis (hydroxymethyl) phosphonium chloride (THPC), gold (III) chloride trihydrate (HAuCl₄), ascorbic acid (AA), polyvinylpyrrolidone (PVP), PBS buffer pH 7.4 tablet, bicarbonate buffer pH 8.5, Tween 20, 2-phospho-L-ascorbic acid, anti-rabbit IgG produced in goat (R2004), IgG from rabbit serum (I5006), anti-rabbit IgG-conjugated alkaline phosphatase antibody produced in goat (A3687), and anti-PSA were purchased from Sigma-Aldrich (St. Louis, MO, USA) and used without further purification. Ethyl alcohol (EtOH) and aqueous ammonium hydroxide (NH₄OH, 27%) were purchased from Daejung (Siheung, Korea). F96 Maxisorp Nunc-Immuno microplate was purchased from Thermo Fisher Scientific (Roskilde, Denmark).

Pham X.H., Hahm E., Kim T.H., Kim H.M., Lee S.H., Lee Y.S., Jeong D.H, & Jun B.H. (2018). Enzyme-catalyzed Ag Growth on Au Nanoparticle-assembled Structure for Highly Sensitive Colorimetric Immunoassay. Scientific Reports, 8, 6290.

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Neuronal Differentiation of hiNPCs

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hiNPCs were plated onto Geltrex-coated coverslips and supplemented with a neuronal differentiation medium, which was comprised of DMEM/F-12 (Thermo Fisher Scientific) medium supplemented with B27 without Vitamin A, 50 μg/ml 2-phospho-L-ascorbic acid (Sigma-Aldrich), 20 ng/ml BDNF and GDNF (Peprotech), and 0.5 mM dbcAMP (Enzo life science, USA). Half of the volume of total media was replaced every other day.

Lee M., Sim H., Ahn H., Ha J., Baek A., Jeon Y.J., Son M.Y, & Kim J. (2019). Direct Reprogramming to Human Induced Neuronal Progenitors from Fibroblasts of Familial and Sporadic Parkinson’s Disease Patients. International Journal of Stem Cells, 12(3), 474-483.

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Isolation and Culture of Dermal Fibroblasts from Wild-type and Classic EDS Mice

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Independent dermal fibroblast strains were isolated from 20 day old mice of Col5a1^+/+ (wild-type) and Col5a1^+/- (classic EDS) and were utilized at passages 2-6. All experiments were repeated using at least 3 different Col5a1^+/+ and Col5a1^+/- strains. All strains from a given genotype produced comparable results. Differences between strain or passage number were comparable to the differences seen between replicate experiments. Briefly, subscapular dermis (∼2cm × 2cm) was harvested from mice that were cleaned and shaved. Dermal tissue strips were incubated in trypsin at 37°C for 20 min with constant agitation and the epidermis was peeled off. Tissue strips were minced and incubated in 2.5mg/ml collagenase B (Roche 11088807001) in DMEM with HEPES at 150rpm in 37°C shaker for 1.5-2 hours until tissue pieces were mostly digested. The digested tissues were filtered through a cell strainer, and fibroblasts were collected and washed two times with DMEM (Invitrogen, 11995) supplemented with 10% fetal bovine serum (FBS). Fibroblasts were cultured at 37°C in DMEM with or without 10% FBS and 1mM 2-phospho-L-ascorbic acid (Sigma, 49752) according to experiments.

DeNigris J., Yao Q., Birk E.K, & Birk D.E. (2015). Altered dermal fibroblast behavior in a collagen V haploinsufficient murine model of classic Ehlers-Danlos syndrome. Connective tissue research, 57(1), 1-9.

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Osteogenic Differentiation of MSCs

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Induced osteogenic differentiation of MSCs was performed according to the accepted protocol of culturing cells in DMEM-advanced medium containing 10⁻⁸ M dexamethasone, 10 mM beta-glycerol 2-phosphate and 0.2 mM 2-phospho-L-ascorbic acid (Sigma-Aldrich; Merck KGaA) (18 (link)).

Xue L., Li Y, & Chen J. (2017). Duration of simulated microgravity affects the differentiation of mesenchymal stem cells. Molecular Medicine Reports, 15(5), 3011-3018.

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Osteogenic Differentiation of hTMSCs

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hTMSCs were cultured in a 24-well plate until cells were 100% confluent. Then, cells were treated with osteogenic medium, the solution of 10 mM glycerol-2-phosphate (Sigma), 1% of 2-Phospho-L-ascorbic acid (Sigma), 1% dexamethasone (Sigma), 10% fetal bovine serum (Biowest), and 1% penicillin/streptomycin (10,000 U/mL) (Gibco) in Dulbecco’s modified Eagle’s medium. For bioglass-treated groups, osteogenic medium with 0.5%, 1.5%, and 2.5% bioglass concentration was used. The culture medium was changed every other day.

Kwon S., Lee S.S., Sivashanmugam A., Kwon J., Kim S.H., Noh M.Y., Kwon S.K., Jayakumar R, & Hwang N.S. (2018). Bioglass-Incorporated Methacrylated Gelatin Cryogel for Regeneration of Bone Defects. Polymers, 10(8), 914.

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