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5 protocols using t butyl acrylate

1

Biomimetic Stem Cell Microenvironment

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t-Butyl acrylate, AIBN, ethyl acetate, methanol, polyacrylic acid (PAA), hexafluoroisopropanol paraformaldehyde, Triton-100, β-glycerophosphate, dexamethasone and ascorbic acid were purchased from Sigma and used as received without further purification. Gelatin (from porcine skin) were purchased from Milliporesigma. Collagen IV (mice) were purchased from Corning. N-Benzophenone acrylate (BPA) and BM1432 (RAFT agent) were donated from Paul Pasic. FITC–RGD with amine as the end group was purchased from BioMATIK (Lot#: P130801, Taiwan). Dulbecco's modified Eagle medium (DMEM), fetal bovine serum (FBS), MEM-a medium (PM150421), penicillin and streptomycin solution were all purchased from Gibico. CCK-8 kits were purchased from Dojindo Molecular Technologies. cRGD were purchased from Qyaobio, China. Fibronectin was purchased from Stem Cell Technology.
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2

Synthesis of Functional Nanomaterials via RAFT Polymerization

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Poly(ethylene glycol) methyl ether acrylate (PEGA, average Mn = 480 g mol -1 ), n-butyl acrylate (n-BA, >99%) and t-butyl acrylate (t-BA 98%), Bromo-propionic acid (>99%), 1butanethiol (99%), carbon disulphide (>99%) and tannic acid (ACS reagent) were obtained from Sigma-Aldrich. All monomers above were passed through basic aluminium oxide to remove inhibitor before use. Chloroform-d3 (99.8% D atom), was obtained from Sigma Aldrich and used for 1 H NMR spectroscopy. Thermal initiator 4,4′-azobis(4-cyanovaleric acid) (ACVA, >98%, Aldrich), was used as received. Cyanine 7.5 amine was purchased from Lumiprobe. TEM grids were purchased from EM Resolutions Ltd (Sheffield, UK). RAFT agent, 2-(((butylthio)carbonothioyl)thio)propanoic acid (PABTC) was synthesised as previously described. 22
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3

Synthesis and Characterization of Polystyrene Films

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Styrene (S) (99%, Sigma-Aldrich, Schnelldorf, Germany) and t‒butyl acrylate (tBA) (98%) (Sigma-Aldrich, Schnelldorf, Germany) were distilled under reduced pressure over calcium hydride prior to their use. Polystyrene (PS) (measured by gel permeation chromatography (GPC), weight-averaged molecular weight Mw = 3 × 106 g mol−1; polydispersity index (PDI) = 1.80) was purchased from Polysciences (USA). Copper (I) bromide (CuBr) (98%), N,N,N′,N′′,N′′‒pentamethyldiethylenetriamine (PMDETA) (99%), and ethyl-2-bromoisobutyrate (EtBr) (98%) acquired from Sigma-Aldrich (Schnelldorf, Germany) and tetrahydrofuran (THF) (HPLC grade) from Scharlab (Barcelona, Spain) were used without further purification. Polymer solutions used for formation of the films were cast in round glass coverslips, 20 mm diameter, purchased from Marienfeld (Lauda-Königshofen, Germany). MilliQ-grade water was used in these studies.
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4

Atom Transfer Radical Polymerization of Styrene and t-Butyl Acrylate

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Styrene (St) (Sigma, Aldrich, 99 %) and t-butyl acrylate (tBA) (Sigma-Aldrich, 98 %) were distilled under reduced pressure over calcium hydride prior to use. Copper (I) bromide (CuBr) (Sigma-Aldrich, 98%), 2,2'-bipyridyl (bipy) (Sigma-Aldrich, 99 %), N,N,N',N'',N'',pentamethyldiethylenetriamine (PMDETA) (Sigma-Aldrich, 99 %), phenylethyl bromide (PhEBr) (Sigma-Aldrich, 97 %), and other solvents were used as received. The diblock copolymers were prepared by atom-transfer radical polymerization (ATRP) in two steps following previously reported procedures [39] .
Polystyrene filaments were purchased from Impresoras 3D (ESUN HIPS). According to the supplier contains around 85 wt% of PS, 10 wt% acrylonitrile and 5 wt% butadiene. High molecular weight polystyrene (Aldrich, Mw =2.50•10 5 g/mol) was used as polymeric matrix. Chloroform (CHCl3) was purchased from Scharlau.
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5

Synthesis of Polystyrene-b-Poly(Acrylic Acid) Diblock Copolymers

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The monomers, t-butyl acrylate (tBA) (Sigma-Aldrich, 98 %) and styrene (St) (Sigma, Aldrich, 99 %), required purification by first dried over calcium hydride and distilled using vacuum techniques. The following list of compounds were used as received: 2,2'-bipyridyl (bipy) (Sigma-Aldrich, 99 %), phenylethyl bromide (PhEBr) (Sigma-Aldrich, 97 %), N,N,N',N'',N'',pentamethyldiethylenetriamine (PMDETA) (Sigma-Aldrich, 99 %), Copper (I) bromide (CuBr) (Sigma-Aldrich, 98%) and other solvents. Coupling reactions were carried out using N-Hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (Sigma-Aldrich -USA). The polystyrene-b-poly(acrylic acid) diblock copolymers were prepared following the procedures already reported by our group [39] .
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