ࡱ> r  !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqstuvwxRoot Entry F"*SummaryInformation(DocumentSummaryInformation8WordDocument8  !"#$%&'()*+,-./0123456789:;<=>?@ABCDEOh+'0 ( 4 @LT\d AsposeNormalchenlin1@pk@*Microsoft Office Word՜.+,D՜.+,\   (\dlKSOProductBuildVer2052-9.1.0.51840Table Data  PKSKS8 \^|H ^ r ~ ? $hF @ ac 0  NwmTupuiryb gPlQS H9c2 'Y ڀ΀_~ހ General Information: Organism: Rattus norvegicus, rat heart/myocardium adherent Tissue: Culture Properties: Morphology: myoblast Culture Method: Complete Growth Medium: DMEM (Gibco 12800-017)+10% FBS+ Penicillin/Streptomycin Volumes are given for a 25cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. 1. 2. Remove and discard culture medium. Briefly rinse the cell layer with 0.25%(w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Subculturing: 3. Add 1.0 to 2.0mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed. 4. 5. 6. Add 6.0 to 8.0mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37C, 5% CO2. Subcultivation Ratio: Cryopreservation: A subcultivation ratio of 1:2 to 1:4 is recommended Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase  laNy 1. 2. 6e0R~ހTHQ‰[~ހt/f&T[}Y W{Qm/f&T gom0QmJmI{sa g Nsa SuSeTbNT|0 N~~ހffN N~ހvsQOo` Y~ހb_`0@b(uW{QW0@nkO0@b ~ހVP[I{ nxO~ހW{QagNN01uNW{QagN NN [~ހQs # N1u[7bLbb0 3. (u 75%R|db~ހthb >f_\ N‰[~ހr`0VЏ4X~ހO g\ϑ NtX1= \~ހnNW{Q{QYnW{QǏY )YQSQ‰[0dkeYpe~ހGWO 4X ~ހN N4X(uSv݄grKm[~ހ;mR Yg[~ހ;mRck8^ \ ~ހy_T(ueW{QWQ!k4XW{QYggr~g>f:y~ހe;mR b NgqGrS eTbNT| Oo`nxTbN:N`QMQ9[N!k0 4. 5. 6. 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