The choline chloride has a melting point of 302 °C and a purity of 97.0% (bought from Iolitech) where β-alanine has a melting point of 258 °C and a purity ≥98%, and betaine has a melting point of 310 °C, purity ≥98%. DL-lactic acid with 85% purity and a melting point of 17 °C was acquired from Sigma Aldrich (St. Louis, MO, USA). All chemicals were used without further purification or treatment. However, the solid choline chloride in the solid state was dried under vacuum at 60 °C for about two days using a desiccator to avoid any humidity absorption. The prepared NADES were stored in sealed containers of 40 mL. The components for this set of NADES systems are summarized in Table 1 .
Dl lactic acid
Manufactured by Merck Group
Sourced in United States, Poland, Germany
DL-lactic acid is a chemical compound used in various industrial and scientific applications. It is a racemic mixture of the D-lactic acid and L-lactic acid enantiomers. DL-lactic acid is commonly used as a pH regulator, preservative, and building block in the synthesis of other chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
D glucose, by Merck Group (8 mentions)
Pyruvic acid, by Merck Group (8 mentions)
Ascorbic acid, by Merck Group (7 mentions)
Progesterone, by Merck Group (7 mentions)
Bovine serum albumin (bsa), by Merck Group (6 mentions)
N2 supplement, by Thermo Fisher Scientific (6 mentions)
Estradiol, by Merck Group (5 mentions)
Choline chloride, by Merck Group (5 mentions)
Fibroblast growth factor (fgf), by Merck Group (4 mentions)
Mercaptoethanol, by Thermo Fisher Scientific (4 mentions)
Stempro supplement, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Stempro 34 sfm, by Thermo Fisher Scientific (4 mentions)
L glutamine, by Thermo Fisher Scientific (4 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (4 mentions)
D biotin, by Merck Group (3 mentions)
Low molecular weight chitosan, by Merck Group (3 mentions)
Glycerol, by Merck Group (3 mentions)
Epidermal growth factor (egf), by Merck Group (3 mentions)
β estradiol, by Merck Group (3 mentions)
38 protocols using dl lactic acid
1
Preparation and Characterization of NADES
2
Optimized Culture Media for Stem Cell Growth
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The culture medium consisted of StemPro‐34 SFM with Stem Pro supplement (Kanatsu‐Shinohara, Ogonuki, et al., 2003 ; Wu et al., 2015 ; Zhang et al., 2014 ; Gibco, #10639–011), 25 μg/mL insulin (Sigma, #I1882), 100 μg/mL transferrin (Sigma, #T1428), 60 μM putrescine (Sigma, #P5780), 30 nM sodium selenite (Sigma, #S5261), 6 mg/mL D‐(+)‐glucose (Sigma, #G7021), 30 μg/mL pyruvic acid (Sigma, #P4562), 1 μL/mL DL‐lactic acid (Sigma, #L7900), 5 mg/mL bovine serum albumin (Calbiochem, #126609), 2 mM L‐glutamine (Millipore, #TMS‐002‐C), 100x β‐mercaptoethanol (Specialty Media, #ES‐007‐E), minimal essential medium vitamin solution (Gibco, #11120–052), 100x nonessential amino acid solution (Gibco, #11140–035), 1% penicillin/streptomycin (Specialty Media, #TMS‐AB2‐C), 0.1 mM ascorbic acid (Sigma, #A4034), 10 μg/mL d‐biotin (Sigma, #B4639), 20 ng/mL recombinant human epidermal growth factor (Gibco, #PMG8041), 10 ng/mL recombinant human basic fibroblast growth factor (PeproTech, #AF‐100‐18B‐250), 10 ng/mL recombinant human GDNF (PeproTech, #450–10‐250), 1 IU/mL Leukemia Inhibitory Factor (Millipore, #ESG1107), and 1% fetal bovine serum (HyClone, #SH30071.03). Cells were maintained at 37°C with 5% CO2.
3
Biocompatible Chitosan-Alginate Hydrogels
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All reagents used in this experiment were purchased from Sigma-Aldrich (Poznan, Poland), unless indicated otherwise.
All experimental procedures were approved by the II Local Ethics Committee of Environmental and Life Science University (Dec. No. 177/2010 of 15 December 2010) and the Local Bioethics Committee of Wroclaw Medical School (registry number KB-177/2014).
Low molecular weight chitosan (DD = 75%–85%; Mw = 150 × 103) and DL lactic acid (85% syrup) were obtained from Sigma Aldrich, Poznan, Poland. Alginate FD 901 AR was purchased from Danisco GRINDSTED®, Grindsted, Denmark. Lysozyme from white egg hen with 2000 U/mg activity was obtained from Ovopol, Nowa Sol, Poland.
All experimental procedures were approved by the II Local Ethics Committee of Environmental and Life Science University (Dec. No. 177/2010 of 15 December 2010) and the Local Bioethics Committee of Wroclaw Medical School (registry number KB-177/2014).
Low molecular weight chitosan (DD = 75%–85%; Mw = 150 × 103) and DL lactic acid (85% syrup) were obtained from Sigma Aldrich, Poznan, Poland. Alginate FD 901 AR was purchased from Danisco GRINDSTED®, Grindsted, Denmark. Lysozyme from white egg hen with 2000 U/mg activity was obtained from Ovopol, Nowa Sol, Poland.
4
Chitosan-Based Hydrogel Preparation
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Low-molecular-weight chitosan (DD = 75%–85%), dl -lactic acid (85% syrup) and 99% glycerol were obtained from Sigma Aldrich, Poznań, Poland. Hydroxypropyl methylcellulose, Methocel™, was purchased from Dow Chemical Co., Midland, MI, USA. Cellulase CP CONC (C) with an activity of 120 U/mg and side activity (typical) of 30 U/mg of β- glucanase was produced by the fermentation of non-GMO Trichoderma longibrachiatum (formerly Trichoderma reesei) that was obtained from Dyadic (Jupiter, FL, USA).
5
Isolation and Culture of Mouse SSCs
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Mouse SSCs were isolated and cultured from 6-day-old male F1 progeny of DBA/2 × CAG-EGFP mice, as previously described [27 (link)]. In brief, SSCs (> 20 passages) were cultured on mitomycin C (MMC)-treated mouse embryonic fibroblast (MEF) feeder cells with SSC culture medium. The SSC culture medium included StemPro-34 SFM supplemented with StemPro supplement (Invitrogen, Carlsbad, CA, USA), human basic fibroblast growth factor (100 μg/ml transferrin, 25 μg/ml insulin, 10 ng/ml; Invitrogen), recombinant human epidermal growth factor (20 ng/ml; Invitrogen), recombinant human glial cell line-derived neurotrophic factor (10 ng/ml; Invitrogen), d -(+)-glucose (6 mg/ml; Invitrogen), putrescine (60 mM; Invitrogen), sodium selenite (30 nM; Invitrogen), l -glutamine (2 mM; Invitrogen), pyruvic acid (30 μg/ml), dl -lactic acid (1 μl/ml; Sigma, St. Louis, MO, USA), bovine serum albumin (5 mg/ml; Sigma), 2-mercaptoethanol (10 μM; Sigma), 1× MEM vitamin solution (Invitrogen), 1× nonessential amino acid solution (Invitrogen), ascorbic acid (0.1 mM), d -biotin (10 μg/ml; Sigma), 1% fetal bovine serum (Gibco), and 1× penicillin/streptomycin solution (Invitrogen). The medium was changed every 2–3 days.
6
Synthesis and Characterization of Polyurethane-Based Composites
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PTHF [Mw (weight-average molecular weight) = 1000 g/mol, 98%] and POE (Mw = 1000 g/mol, 98%) were purchased from Sigma-Aldrich and degassed at 130°C for 3 hours before use. TMP (Mw = 134.17 g/mol, 97%), DBTDL (95%), HEMA (Mw = 130.14 g/mol, ≥99 DBTDL (Mw = 631.56 g/mol, 95%), acetone (anhydrous ≥99.5%), sodium chloride, dl -lactic acid (~90%), potassium chloride, sodium hydroxide, and urea were also received from Sigma-Aldrich and used without any purification. IPDI (a mixture of isomers, 98%), methyl ethyl ketone (MEK; ≥99%), 1-hydroxycyclohexyl phenyl ketone (IRGACURE 184; Mw = 204.3 g/mol, 99%), Ag flakes (10 μm), the SEBS (Tuftec H1052) with 20/80 S/EB weight ratio, and the hydrophilic textile were obtained from Alfa Aesar, Fisher Chemical, Ciba Specialty Chemicals, Puwei Applied Materials Technology, Asahi Kasei Corporation, and MHTC Technology Company, respectively. Deuterated solvents for NMR characterization were obtained from Cambridge Isotope Laboratories Inc. Deionized water was used throughout the study.
7
Germ Stem Cell Culture Protocol
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Then SSCs that digested testicular cells were cultured in Germ Stem Cells (GSCs) culture media at 37°C with a 5% CO2 supplementation. The GSC culture media was composed of Stem-Pro-34 medium, 1% N2-supplement (Invitrogen, USA), 5 μg/ml of bovine serum albumin (Sigma Aldrich, USA), 1% L-glutamine (PAA laboratories, USA), 1% penicillin/streptomycin (PAA laboratories, USA), 20 ng/ml of epidermal growth factor (EGF) (Sigma Aldrich, USA), 30 ng/ml of estradiol (Sigma Aldrich, USA), 6 mg/ml of D-glucose (Sigma Aldrich, USA), 0.1% beta mercaptoethanol (Invitrogen, USA), 100 μg/ml of ascorbic acid (Sigma Aldrich, USA), 1% nonessential amino acids (PAA laboratories, USA), 30 μg/ml of pyruvic acid (Sigma Aldrich, USA), 60 ng/ml of progesterone (Sigma Aldrich, USA), 1% MEM vitamins (PAA laboratories, USA), 100 U/ ml of human LIF (MilliporeSigma, USA), 10 ng/ ml of FGF (Sigma Aldrich, USA), 8 ng/ml of GDNF (Sigma Aldrich, USA), 1% ES cell qualified FBS, and 1 μl/ml of DL-lactic acid (Sigma Aldrich, USA) (27 (link), 28 (link)).
8
Analytical Quantification of Metabolites
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N-acetyl-l-aspartic acid (NAA, Sigma-Aldrich, 00920, CAS: 997-55-7, Scheme 1), DL-lactic acid (Sigma-Aldrich, 69785, CAS: 50-21-5), l-alanine (Sigma-Aldrich, A7469, CAS: 56-41-7), creatine monohydrate (Sigma-Aldrich, C3630, CAS: 6020-87-7), choline chloride (Sigma-Aldrich, C7017, CAS: 67-48-1), l-glutamic acid (Sigma-Aldrich, 49449, CAS: 56-86-0), myo-Inositol (Sigma-Aldrich, I7508, CAS: 87-89-8) were purchased and used without further purification.
9
GDNF-Supplemented Mouse Embryonic Stem Cell Culture
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GDNF-mESCCM consisted of high-glucose DMEM supplemented with 15% (v/v) heat-inactivated FBS, 0.1 mM β-mercaptoethanol (Gibco Invitrogen, Grand Island, NY, USA), 1% (v/v) non-essential amino acid (NEAA; Gibco Invitrogen, USA), 2 mM l -glutamine (Gibco Invitrogen, USA), 1% (v/v) antibiotic–antimycotic solution, 1,000 U/mL mouse leukemia inhibitory factor (mLIF; Chemicon International, Inc., Temecula, CA, USA), and 10 ng/mL GDNF (R&D Systems, Inc., Minneapolis, MN, USA). MEM alpha medium (Gibco Invitrogen, USA), which served as pSSCCM, was supplemented with 1% (v/v) antibiotic–antimycotic solution, 1% (v/v) NEAA, 0.1 mM β-mercaptoethanol, N2-1 supplement (Merck Millipore; Darmstadt, Germany), DL-lactic acid (Sigma-Aldrich, USA), 1% (v/v) MEM vitamin solution (Sigma-Aldrich, USA), 30 ng/mL β-estradiol (Sigma-Aldrich, USA), 1% (v/v) heat-inactivated FBS, 10 ng/mL basic fibroblast growth factor (bFGF; PeproTech, Inc., Rocky Hill, NJ, USA), 20 ng/mL epidermal growth factor (EGF; PeproTech, Inc., USA), 1,000 U/mL mLIF, and 10 ng/mL GDNF.
10
Synthesis and Modification of Nanoparticles
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Chemicals for the nanoparticle synthesis were purchased and used without further purification: ethanol absolut (VWR), diethylene glycol (99%, Alfa Aesar), N-methyl-2,2‘-iminodiethanol (Merck), sodium hydroxide pellets (Merck), Hydrochloric acid solution (in water 1 M, Grüssing GmbH), Sodium hydroxide solution (in water 1 M, Grüssing GmbH), acetone (technical grade, Carl Roth GmbH), iron(II) chloride tetrahydrate (Glentham Life Sciences) and iron(III) chloride hexahydrate (puriss. p.a. reag., ≥99%, Sigma-Aldrich). Ligands for the nanoparticle modification were obtained from Sigma-Aldrich L-histidine monohydrochloride monohydrate (≥99%), L-lysine monohydrochloride (BioUltra, ≥99.5% (AT)); 4-mercaptobenzoic acid (99%); 4-aminobenzoic acid (Reagent Plus, ≥ 99%); DL-Lactic Acid (~90%), phosphacholine chloride calcium salt tetrahydrate (Sigma grade); cysteamin (> 98%); L-(+)-tartaric acid (99%), from Alfa Aesar L-Arginine (98%), L-(−)-malic acid (99%), 4-hydroxybenzoic acid (99%), (±) Mandelic Acid (99%) and from Grüssing GmbH trisodium citrate dihydrate.
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