Human bone marrow MSC were obtained from five donors (41–65 years old) undergoing treatment with intra-articular injections of autologous MSC at Avenue Cell Clinic in accordance with a protocol approved by the Institutional Ethics Committee. Human MSC around passage 4–5 were induced to undergo chondrogenic differentiation. For the pellet culture, 2 × 105 cells were placed in 96 deep well polypropylene plates and centrifuged at 500 × g for 5 minutes with basal media; DMEM/HG supplemented with 1% ITS premix (Corning; Corning, NY), 50 µg/mL 2-Phospho-L-ascorbic acid trisodium, 40 µg/mL L-proline (Sigma-Aldrich), 1% PS. The next day, the media was changed to chondrogenic-induction media; basal media supplemented with 10 nM dexamethasone (Wako), 10 ng/mL recombinant human transforming growth factor-β1 (rhTGF-β) (Oriental Yeast Co) and 100 ng/mL recombinant human bone morphogenetic protein-2 (rhBMP-2) in the presence or absence of KAG-308. The media and KAG-308 were changed twice a week. After 21 days, the pellets were analyzed.
2 phospho l ascorbic acid trisodium
Manufactured by Merck Group
Sourced in Senegal
2-Phospho-L-ascorbic acid trisodium is a chemical compound used in laboratory settings. It is a derivative of the antioxidant vitamin C (ascorbic acid), with a phosphate group attached at the 2-position. The core function of this compound is to serve as a reagent or intermediate in various biochemical and analytical procedures.
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Lab products found in correlation
L proline, by Merck Group (2 mentions)
Dexamethasone (dex), by Fujifilm (1 mentions)
Its premix, by Corning (1 mentions)
Poly ε caprolactone, by Merck Group (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Pentobarbital, by Merck Group (1 mentions)
Hexafluoroisopropanol hfip, by Maclin Biochemical Technology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Nitric acid, by Merck Group (1 mentions)
Multi walled carbon nanotubes mwcnt, by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Sodium dodecyl sulphate (sds), by Merck Group (1 mentions)
Graphene oxide, by Merck Group (1 mentions)
Magnesium chloride hexahydrate, by Merck Group (1 mentions)
Lithium perchlorate, by Merck Group (1 mentions)
Hydrogen chloride, by Merck Group (1 mentions)
Tris hydroxymethyl aminomethane, by Avantor (1 mentions)
5 protocols using 2 phospho l ascorbic acid trisodium
1
Chondrogenic Differentiation of Human Bone Marrow MSCs
2
Chondrocyte Pellet Culture with KAG-308
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Mouse primary chondrocytes were centrifuged at 500 × g for 5 minutes and cultured as pellets (2 × 105 cells/pellet) in 96 deep well polypropylene plates (Evergreen Scientific, Vernon, CA) with DMEM supplemented with 10% FBS charcoal stripped, 1% PS and 50 µg/mL 2-Phospho-L-ascorbic acid trisodium (Sigma-Aldrich) in the presence or absence of KAG-308, as previously described62 (link). The medium was replaced every other day with KAG-308. Cultured pellets were analyzed 2 weeks after treatment.
3
Tailoring PCL Scaffolds for Cell Culture
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Poly (ε-caprolactone) (PCL, average Mn = 80 000), collagenase type I (collagenase from Clostridium histolyticum for general use), Poly-L-lysine (PLL), paraformaldehyde (PFA), 2-phospho-L-ascorbic acid trisodium, bovine serum albumin (BSA), and pentobarbital were obtained by Sigma-Aldrich (Sigma-Aldrich, Saint Louis, United Kingdom). Hexafluoroisopropanol (HFIP) was purchased from Macklin (Macklin, China).
4
Nanomaterial-Enhanced Enzyme Assay Protocol
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Hydrogen chloride (37%, w/w), nitric acid (68% w/w), lithium perchlorate, magnesium chloride hexahydrate, L-ascorbic acid (AA), 2-phospho-L-ascorbic acid trisodium (AAP), sodium dodecyl sulphate (SDS), Tween 80, Triton X-100, bovine serum albumin (BSA), glutaraldehyde (GA) (25%, w/w) were of reagent grade and were purchased from Sigma-Aldrich. Alkaline phosphatase (ALP) from bovine intestinal mucosa, 37 units/mg solid, was obtained from Sigma Aldrich (Saint Louis, MO, USA). Tris-hydroxymethyl-aminomethane (TRIS) was purchased from VWR chemicals.
Graphene oxide (GO; 4 mg/mL water suspension) and multi-walled carbon nanotubes (MWCNTs; diameter 20–50 nm) were bought from Sigma-Aldrich.
Graphene oxide (GO; 4 mg/mL water suspension) and multi-walled carbon nanotubes (MWCNTs; diameter 20–50 nm) were bought from Sigma-Aldrich.
5
Primary Hepatocytes Isolation and Culture
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Primary human hepatocytes (PHHs) were collected from humanized liver chimeric NOG‐TKm30 mice[19 ] using the two‐step collagenase–pronase liver perfusion method as previously reported.[20 ] The collected PHHs were seeded in 12‐well or 24‐well collagen type I‐coated microplates (AGC Techno Glass Co., Ltd.). The PHH culture medium was Dulbecco's modified Eagle's medium (DMEM) (Nacalai Tesque, Inc.) supplemented with 5 μg/ml L‐proline (Sigma‐Aldrich), 32.2 μg/ml 2‐phospho‐L‐ascorbic acid trisodium (Sigma‐Aldrich), 0.02 μg/ml dexamethasone (Sigma‐Aldrich), 0.005 μg/ml recombinant human epidermal growth factor (Sigma‐Aldrich), 25 μl/L human insulin solution (Sigma‐Aldrich), 20 mM 4‐(2‐hydroxyethyl)‐1‐piperazine ethanesulfonic acid (Nacalai Tesque, Inc.), 0.37% NaHCO3 (Nacalai Tesque, Inc.), 2% dimethyl sulfoxide (DMSO) (Nacalai Tesque, Inc.), 10% fetal bovine serum (FBS; Life Technologies), 100 units/ml penicillin, 100 μg/ml streptomycin, and 0.25 μg/ml amphotericin B (Life Technologies). The culture medium was changed on the indicated days. HepG2‐hNTCP‐C4‐iCas9 cells were maintained in DMEM/F‐12 (Life Technologies) containing G418 Disulfate Aqueous Solution and 10% FBS at 37°C under 5% CO2. Doxycycline (DOX) was administered at 1 μg/ml to induce Cas9.
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