416
Appendix
cium pantothenate 0.24, made up to 1,000 ml with distilled water. The pH of the medium should be adjusted to 6.7 with 1 N KOH and 1 N HCl.
Sengel's medium (Sengel, 1960)
Salt solution (Holtfreter's solution, 1931): NaC13,500 mg, KCI SO mg, CaCl 2 2H 2 0 100 mg,
and NaHC0 3 200 mg, made up to 1,000 ml with distilled water.
Culture medium: 5 parts of 1 % agar in the above salt solution, 4 parts of 0.1 % glucose
in the above salt solution, and 3 parts chicken embryo (9 days old) extract diluted to
50% with the above salt solution.
SH-7 medium (Furuta and Shimozawa, 1983)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
NaCI 2,483.0, KCI 496.0, CaCl 2 6.0, MgCl 2 152.0, NaHC0 3 1,769.0, glucose 50.0, trehalose 50.0, L-a-alanine 41.2, L-arginine hydrochloride 6.1, L-asparagine monohydrate
14.5, L-aspartic acid 3.1, L-cystine 3.3, L-glutamic acid 3.7, L-glutamine 3.7, glycine 3.0,
L-histidine dihydrochloride 4.1, L-isoleucine 3.3, L-Ieucine 2.3, L-Iysine hydrochloride
5.8, L-phenylalanine 5.7, L-serine 11.8, L-threonine 27.2, L-tyrosine 2.8, L-valine 4.9, thymidine 0.35, uridine 22.4, hypoxanthine 2.0, tricine 2,750.0, putrescine 0.15, phenol
red 2.0, D-biotin 0.007, choline chloride 7.0, calcium pantothenate 0.25, folic acid 0.5,
inositol 9 .0, pyridoxal hydrochloride 0.03, riboflavin 0.02, thiamine hydrochloride 0.15,
cyanocobalamin 0.5, nicotinamide 0.02, linoleic acid 0.04, a-lipoic acid 0.1, FBS (dialyzed and inactivated) 100.0 ml, and chicken embryo extract 40.0 ml, made up to
1,000 ml with distilled water.
Shields and Sang's Medium (Shields and Sang, 1970)
The concentrations are given as mg/l,OOO ml culture medium.
MgS04·7H20 5,130, CaCI2·6H20 1,740, KCI3,130, NaC1860, NaH 2 P0 4 ·2H 2 0 880, KHC0 3
180, monosodium malate·2H20 950, monosodium a-ketoglutarate 420, disodium fumarate 80, disodium succinate·6H20 140, monosodium L-glutamate 2,460, L-aspartic
acid ISO, L-threonine 500, L-serine 350, L-asparagine 300, L-glutamine 600, L-proline
400, glycine 500, L-a-alanine 1,650, L-valine 420, L-methionine 120, L-isoleucine 270, Lleucine 400, L-tyrosine 260, L-phenylalanine 240, ~-alanine 100, L-histidine 550, L-tryptophan 100, L-arginine 500, L-Iysine 680, L-cystine 200, L-cysteine 800, glutathione 5,
glucose 4,600, and TC-Yeastolate (Difco Laboratories) 2,000. The pH of the medium is
adjusted to 6.9 with 1 % NaOH. Use the medium by adding 10% FBS.
Shields and Sang's medium modified by Poels (1972)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 860, NaH2P04·2H20 880, KCI 3,130, KHC0 3 180, CaCl 2 ·2H 2 0 1,160, MgS0 4 ·7H 2 0
5,130, monosodium malate·2H 2 0 950, monosodium a-ketoglutarate 420, disodium
succinate·6H 2 0 140, disodium fumarate 80, monosodium pyruvate 20, monosodium Lglutamate 760, L-aspartic acid 90, L-threonine 150, L-serine 175, L-asparagine 150, Lglutamine 300, L-proline 400, glycine 175, L-a-alanine 750, L-valine 90, L-methionine
150, L-isoleucine SO, L-Ieucine 90, L-tyrosine 30, L-phenylalanine 30, ~-alanine 120, Lhistidine 190, L-tryptophan 100, L-arginine 210, L-Iysine 70, L-cystine 10, L-cysteine 60,
TC-Yeastolate 2,000, glucose 300, trehalose 800, and FBS 100 ml. The pH of the medium is adjusted to 6.9 with 1 N NaOH.
Appendix
cium pantothenate 0.24, made up to 1,000 ml with distilled water. The pH of the medium should be adjusted to 6.7 with 1 N KOH and 1 N HCl.
Sengel's medium (Sengel, 1960)
Salt solution (Holtfreter's solution, 1931): NaC13,500 mg, KCI SO mg, CaCl 2 2H 2 0 100 mg,
and NaHC0 3 200 mg, made up to 1,000 ml with distilled water.
Culture medium: 5 parts of 1 % agar in the above salt solution, 4 parts of 0.1 % glucose
in the above salt solution, and 3 parts chicken embryo (9 days old) extract diluted to
50% with the above salt solution.
SH-7 medium (Furuta and Shimozawa, 1983)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
NaCI 2,483.0, KCI 496.0, CaCl 2 6.0, MgCl 2 152.0, NaHC0 3 1,769.0, glucose 50.0, trehalose 50.0, L-a-alanine 41.2, L-arginine hydrochloride 6.1, L-asparagine monohydrate
14.5, L-aspartic acid 3.1, L-cystine 3.3, L-glutamic acid 3.7, L-glutamine 3.7, glycine 3.0,
L-histidine dihydrochloride 4.1, L-isoleucine 3.3, L-Ieucine 2.3, L-Iysine hydrochloride
5.8, L-phenylalanine 5.7, L-serine 11.8, L-threonine 27.2, L-tyrosine 2.8, L-valine 4.9, thymidine 0.35, uridine 22.4, hypoxanthine 2.0, tricine 2,750.0, putrescine 0.15, phenol
red 2.0, D-biotin 0.007, choline chloride 7.0, calcium pantothenate 0.25, folic acid 0.5,
inositol 9 .0, pyridoxal hydrochloride 0.03, riboflavin 0.02, thiamine hydrochloride 0.15,
cyanocobalamin 0.5, nicotinamide 0.02, linoleic acid 0.04, a-lipoic acid 0.1, FBS (dialyzed and inactivated) 100.0 ml, and chicken embryo extract 40.0 ml, made up to
1,000 ml with distilled water.
Shields and Sang's Medium (Shields and Sang, 1970)
The concentrations are given as mg/l,OOO ml culture medium.
MgS04·7H20 5,130, CaCI2·6H20 1,740, KCI3,130, NaC1860, NaH 2 P0 4 ·2H 2 0 880, KHC0 3
180, monosodium malate·2H20 950, monosodium a-ketoglutarate 420, disodium fumarate 80, disodium succinate·6H20 140, monosodium L-glutamate 2,460, L-aspartic
acid ISO, L-threonine 500, L-serine 350, L-asparagine 300, L-glutamine 600, L-proline
400, glycine 500, L-a-alanine 1,650, L-valine 420, L-methionine 120, L-isoleucine 270, Lleucine 400, L-tyrosine 260, L-phenylalanine 240, ~-alanine 100, L-histidine 550, L-tryptophan 100, L-arginine 500, L-Iysine 680, L-cystine 200, L-cysteine 800, glutathione 5,
glucose 4,600, and TC-Yeastolate (Difco Laboratories) 2,000. The pH of the medium is
adjusted to 6.9 with 1 % NaOH. Use the medium by adding 10% FBS.
Shields and Sang's medium modified by Poels (1972)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 860, NaH2P04·2H20 880, KCI 3,130, KHC0 3 180, CaCl 2 ·2H 2 0 1,160, MgS0 4 ·7H 2 0
5,130, monosodium malate·2H 2 0 950, monosodium a-ketoglutarate 420, disodium
succinate·6H 2 0 140, disodium fumarate 80, monosodium pyruvate 20, monosodium Lglutamate 760, L-aspartic acid 90, L-threonine 150, L-serine 175, L-asparagine 150, Lglutamine 300, L-proline 400, glycine 175, L-a-alanine 750, L-valine 90, L-methionine
150, L-isoleucine SO, L-Ieucine 90, L-tyrosine 30, L-phenylalanine 30, ~-alanine 120, Lhistidine 190, L-tryptophan 100, L-arginine 210, L-Iysine 70, L-cystine 10, L-cysteine 60,
TC-Yeastolate 2,000, glucose 300, trehalose 800, and FBS 100 ml. The pH of the medium is adjusted to 6.9 with 1 N NaOH.