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. Author manuscript; available in PMC: 2023 Oct 28.
Published in final edited form as: J Vis Exp. 2022 Oct 28;(188):10.3791/64554. doi: 10.3791/64554

Table-Materials

Name of Material/ Equipment Company Catalog Number Comments/Description
1 mL Luer-Lok syringe sterile, single use, polycarbonate BD 309628
5 mL Luer-Lok syringe sterile, single use, polycarbonate BD 309646
Alexa Fluor 488 C5 maleimide Invitrogen A10254 For synthesis of fluorescently-labeled tetrazine
Alexa Fluor 647 Phalloidin Invitrogen A22287 For staining cell culture samples
Aluminum foil VWR 89107–726
Biopsy punch with plunger, 1.0 mm Integra Miltex 69031–01
Biopsy punch, 4 mm Integra Miltex 33–34
Blunt needle, 23G 0.5”, Non-Sterile, Capped SAI Infusion Technologies B23-50
Bottle-top vacuum filter, 0.22 μm Corning CLS430521
Calcium chloride VWR 1B1110 For microgel washing buffer
Capillary-piston assemblies for positive-displacement pipettes, 1000 μL max. volume Rainin 17008609
Capillary-piston assemblies for positive-displacement pipettes, 25 μL max. volume Rainin 17008605
Capillary-piston assemblies for positive-displacement pipettes, 250 μL max. volume Rainin 17008608
Centrifuge tube, 15 mL CELLTREAT 667015B
Centrifuge tube, 50 mL CELLTREAT 229421
Chloroform, ACS grade, Glass Bottle Stellar Scientific CP-C7304 For synthesis of fluorescently-labeled tetrazine
Corona plasma gun, BD-10A High Frequency Generator ETP 11011
CryoTube Vials, Polypropylene, Internal Thread with Screw Cap Nunc 368632
D1 mouse mesenchymal cells ATCC CRL-12424 Example cell line for culture in MAP gels
DAPI Sigma-Aldrich D9542 For staining cell culture samples
Deuterium oxide, 99.9 atom % D Sigma-Aldrich 151882 For NMR spectroscopy
Dialysis tubing, regenerated cellulose membrane, 12-14 kDa molecular weight cut-off Spectra/Por 132703 For purifying HA-NB and HA-Tet
Diethyl ether VWR BDH1121-4LPC For synthesis of fluorescently-labeled tetrazine
Dimethylformamide Sigma-Aldrich 277056 For synthesis of fluorescently-labeled tetrazine
4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) TCI-Chemicals D2919 For modifying HA
Dithiothreitol (DTT) Thermo Scientific R0861 Non-degradable dithiol linker (substitute for MMP-cleavable peptide)
Dulbecco’s Modified Eagle’s Medium - high glucose, w/ 4500 mg/L glucose, L-glutamine, sodium pyruvate, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture Sigma-Aldrich D6429–500ML For D1 cell culture
EMS Paraformaldehyde, Granular VWR 100504–162 For making 4% PFA
Ethanol absolute (200 proof) KOPTEC 89234–850
Heating Plate Kopf Instruments HP-4M
2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) Sigma-Aldrich H3375
Sodium hyaluronate, 79 kDa average molecular weight, produced in bacteria Streptococcus zooepidemicus, pharmaceutical grade, microbial contamination <100 CFU/g, bacterial endotoxins <0.050 IU/mg Contipro N/A 79 kDa average molecular weight was used for HA-Tet synthesis, but these methods but could be adapted for other molecular weights.
IMARIS Essentials software package Oxford Instruments N/A Microscopy image analysis software
Infusion pump, dual syringe Chemyx N/A
Kimwipe Kimberly-Clark 34120
Laboratory stand with support lab clamp Geyer 212100
Liquid nitrogen Airgas NI 180LT22
Lithium Phenyl(2,4,6-trimethylbenzoyl)phosphinate TCI-Chemicals L0290
Methyltetrazine-PEG4-maleimide Kerafast FCC210 For synthesis of fluorescently-labeled tetrazine
2-(4-Morpholino)ethane Sulfonic Acid (MES) Fisher Scientific BP300–100 For modifying HA
Micro cover glass, 24×60mm No. 1 VWR 48393–106
Mineral oil, heavy Sigma-Aldrich 330760
MMP-cleavable dithiol crosslinker peptide (Ac-GCRDGPQGIWGQDRCG-NH2) GenScript N/A
5-Norbornene-2-methylamine TCI-Chemicals 95–10–3 For HA-NB synthesis
Packing tape Scotch 3M 1426
Parafilm Bemis PM996
PEG(thiol)2 JenKem Technology USA A4001-1 For synthesis of fluorescently-labeled tetrazine
Petri dish, polystyrene, disposable, Dia. x H=150 × 15 mm Corning 351058
Pluronic F-127 Sigma-Aldrich P2443 For washing HMPs
Phosphate buffered saline (PBS) 1x Gibco 10010023
RainX water repellent glass treatment Grainger 465D20
RGD peptide (Ac-RGDSPGERCG-NH2) GenScript N/A
Rubber bands Staples 112417
Sodium chloride Chem-Impex 30070 For dialysis
Span® 80 for synthesis Sigma-Aldrich 1338–43–8
Sylgard 184 Silicone Elastomer Electron Microscopy Science 4019862
Syringe filter, Whatman Uniflo, 0.2 μm PES, 13 mm diameter Cytvia 09–928–066
Tetraview LCD digital microscope Celestron 44347
Tetrazine-amine HCl salt Chem-Impex 35098 For HA-Tet synthesis
Triethylamine Sigma-Aldrich 471283 For synthesis of fluorescently-labeled tetrazine
Tris(2-carboxyethyl)phosphine (TCEP) Millipore Sigma 51805–45–9
Triton X-100 VWR 97063–864
Trypan blue solution, 0.4% Thermo Fisher Scientific 15250061
Trypsin EDTA (0.25%), Phenol red Fisher Scientific 25–200–056 For lifting adherent cells to seed in MAP gels
Tygon® formula E-3603 laboratory tubing Millipore Sigma Z765244
UV curing system controller, LX500 LED OmniCure 010–00369R
UV curing head, LED spot UV OmniCure N/A
UV light meter, Traceable VWR 61161–386
Vacuum dessicator Bel-Art 08–594–15C
X-Acto Z Series Precision Utility Knife Elmer’s XZ3601W