1 (1) (2) ( ) (3) (4) (5) (1) (2) (3) 2 (1) ( ) (2) (3) (4) 1 2 1
3 4 5 1 1-20 4 2, 260mOsm/kg 320 mosm/kg HEPES ph 7.2-7.4 ph ph (7.4-7.7), ph (7.0-7.4) NaHCO 3 CO 2 CO 2 CO 2 5 2
NaHCO 3 1.97g/L CO 2 10 NaHCO 3 3.95g/L HEPES ph 7.2-7.4 HEPES 0.34g/L HEPES ph DMEM 2-2-Mercaptoethanol 2-Me 2-Me 2-Me DNA (PHA) 2-Me 78.13 2-Me 1.110-1.120(Do20) 5 10-5 M 0.1M 0.5ml 4 37 12 L- L- 4 36 3
1-20 70 4 1 10 2 3-20-70 4 1-20 37 4 56, 30 complement,,,, 5 37 6 3000rpm, 5 1 2 1 2 4
3 4 5 3 1 500ml 250ml 100ml 2 1cm 200ml 100ml 50ml 25ml 10ml 3 30mm 60mm 120mm 4 1ml 10ml 5 50ml 30ml 15ml 10ml 5ml 6 4 36.5 ±0.5 39 39-40 1 40-41 1 41-42 1 43 1 0 25 35 4 37 80 196 5 95% 5% 5
ph ph 7.2-7.4 ph 7.4-7.6 ph 1 30-60 70% 10 70% 10 2 70% 3 45 4 DMSO TPA 5 CO 2 CO 2 CO 2 CO 2 HEPA 300 / 3000 /HEPA (latent phase),.,.,,.,, 10-24 6
,, 10-30. 24-96, 6-24 (2) (logarithmic growth phase) Mitotic index, MI 1000 0.1%-0.5% 3 5 3 5 (contact inhibition) piled up Density Inhibition 3 (Stagnate phase) Plateau phase ph 1-2 1 2-3 2 7
3 1 2 3 6 g ml mla 63 1000 200000 B 120 200 1000 C 100 100 100 4 8
3-5 2% NaOH 10-20 1% 30 10-20 2% NaOH 5% 30 1 2.5 2 121, 15, 20 121, 15, 20 170, 4 3 70% 1 4 9
RPMI-1640 DMEM- DMEM- IMDM McCoys 5A M199 F10 PBS Phosphate-Buffered Sallines DPBS Dulbecco s Phosphate-Buffered Sallines g/l CaCl 2 (anhyd.)( ) 0.10 KCl 0.20 KH 2 PO 4 0.20 MgCl 2 6H 2 O 0.10 NaCl 8.00 Na 2 HPO 4 7H 2 O 2.16 Hanks Hanks Balanced Salt Solutions HBSS g/l CaCl 2 (anhyd.)( ) 0.14 KCl 0.40 KH 2 PO 4 0.06 MgCl 2 6H 2 O 0.10 10
MgSO 4 7H 2 O 0.10 NaCl 8.00 NaCO 3 0.35 Na 2 HPO 4 0.048 D-Glucose 1.00 Phenol Red 0.01 D-Hanks (D-Hanks Balanced Salt Solutions, D-HBSS) g/l KCl 0.40 KH 2 PO 4 0.06 NaCl 8.00 NaCO 3 0.35 Na 2 HPO 4 0.048 D-Glucose 1.00 Phenol Red 0.01 (EDTA) 1 ph8.0 37 Ca 2+ Mg 2+ D-Hanks 0.25% 1 D-Hanks ph7.2 D-Hanks 4 2 0.25% 0.125% ph 7.2-20 EDTA 4Na EDTA 0.02% 0.25% 1 1 EDTA Hanks EDTA EDTA D-HBSS 4 11
EDTA 4Na 0.05%, 0.53mM EDTA 4Na 0.5g 0.2gEDTA 4Na 1L D-HBSS -20 ph 1 NaHCO3 7.5% 4 ph 10 CO2 2 HEPES 238.31 HEPES 10-50mM, 1M 200ml 47.6 HEPES 1N NaOH ph 7.5-8.0; 4 ph ph ph D- 1 2 3 4 5 (6) 1 4 30 60-20 30-80 16 18-1 -3-80 2 90 3 1 10 7 5 10 7 /ml 4 20 5 DMSO( ) 5 10 DMSO HL-60 DMSO HL-60 glycele, 1-20 1-20 -80 2 DMSO DMSO 12
3 DMSO DMSO 4 DMSO DMSO 1 10 DMSO 20 2 10 20 1 200 400g 5 1 1 37 1 3 2 1ml 25 30ml 24 DMSO 1 37 2 : (1) 37 2 1 2ml 25ml 3 80g 2 3 4 5 3 10 5 /ml 13