Dulbecco s phosphate buffered saline without calcium chloride

N-Isopropylacrylamide (IPAAm) and (3-acrylamidopropyl)trimethylammonium chloride (APTAC) were provided by KJ Chemicals (Tokyo, Japan). IPAAm was recrystallized from hexane, and APTAC was used as received. 3-Aminopropyltriethoxysilane (APTES) was purchased from Shin-Etsu Chemical (Tokyo, Japan). 4-Cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanoic acid (CDPA) as well as Dulbecco’s phosphate buffered saline without calcium chloride and magnesium chloride (DPBS) were obtained from Sigma-Aldrich (St. Louis, Missouri, USA). N-Hydroxysuccinimide (NHS), N,N’-dicyclohexylcarbodiimide (DCC), 4,4ʹ-azobis(4-cyanovaleric acid) (V-501), tris(2-carboxyethyl)phosphine hydrochloride (TCEP), diethyl ether, dehydrated toluene, methanol, acetone, dehydrated dichloromethane (DCM), and 1,4-dioxane were obtained from FUJIFILM Wako Pure Chemicals (Osaka, Japan) and used without further purification. Glass coverslips (24 mm × 50 mm, thickness: 0.2 mm) were purchased from Matsunami Glass (Osaka, Japan). Water used in this study was purified using a Milli-Q Synthesis A10 system (Millipore, Billerica, Massachusetts, USA).

All plastic consumables including cell culture flasks and plates, tubes, and filter tips were purchased from Sarstedt AG & Co. (Germany). Chemical reagents were purchased as follows: Dulbecco’s Phosphate Buffered Saline without calcium chloride and magnesium chloride (#D6662-10X1L, Sigma-Aldrich, Germany); Opti-MEM 1X + GlutaMAX, Reduced serum medium (#1854076, Gibco, Life Technologies, Germany); DMEM High Glucose (4.5 g/l) w/o l-glutamine (#DMEM-HXA, Capricorn Scientific GmbH, Germany); l-glutamine for cell culture (#A3704,0100, Applichem, Germany); MEM nonessential amino acids solution (100x) (#NEAA-B, Capricorn Scientific GmbH, Germany); 2-mercaptoethanol (#BCBS5481, Sigma-Aldrich, Germany); penicillin/streptomycin solution (100x) (#PS-B, Capricorn Scientific GmbH, Germany); Trypsin–EDTA (10×) (#L11-003, GE Healthcare, PPA Laboratories GmbH, Austria); fetal bovine serum (#10270-106, Gibco, ThermoFisherScientific, Germany); dimethyl sulfoxide (#D4540-500ML, Sigma-Aldrich, Germany); LIF (hBA-FL) (#sc-4377, Santa Cruz Biotechnology, Germany); sodium pyruvate (#P2256, Sigma-Aldrich, Germany); gelatin from bovine skin (#G939-100G, Sigma-Aldrich, Germany); and Hoechst 33354 (#62249, Thermo Scientific, Germany). We used a Gene Pulser Xcell system with CE Module from BIO-RAD (Germany) and 4 mm electroporation cuvettes (#748052, Biozym, Germany).