Composition of Salt Solutions and Culture Media
407
200 ml, and dihydrostreptomycin sulfate 100, made up to 1,000 ml with distilled water.
The pH of the medium is adjusted to 6.2 with KOH.
Dulbecco's modified Eagle's medium (Morton, 1970)
The concentrations are given as mg/l,OOO ml medium.
CaClz 200, Fe(N03)3·9HzO 0.1, KCI 400, MgS04 97.67, NaCI 6,400, NaHC0 3 3,700,
NaH z P0 4 ·H z O 125, glucose 1,000, phenol red 15, sodium pyruvate 110, L-arginine hydrochloride 84, L-cystine dihydrochloride 62.57, L-glutamine 584, glycine 30, L-histidine hydrochloride monohydrate 42, L-isoleucine 104.8, L-Ieucine 104.8, L-Iysine hydrochloride 146.2, L-methionine 30, L-phenylalanine 66, L-serine 42, L-threonine 95.2,
tryptophan 16, L-tyrosine 2Na·2HzO 103.79, L-valine 93.6, o-calcium pantothenate 4.0,
choline chloride 4.0, folic acid 4.0, myoinositol 7.0, niacinamide 4.0, pyridoxal hydrochloride 4.0, riboflavin 0.4, and thiamine hydrochloride 4.0.
Durchon and Schaller's medium for Annelida (Durchon and Schaller, 1963)
Mix the following: 7 drops of 1 % agar in sterilized natural seawater containing 0.1 %
glucose, 3 drops of sterilized seawater containing 0.1% glucose, 1 drop of 50% egg
albumin in sterilized seawater containing 0.1 % glucose, 1 drop of horse serum (Difco
Laboratories), 1 drop of 50% chicken embryo extract in sterilized seawater containing
0.1 % glucose, 100 IV of penicillin G, and 0.009 mg of streptomycin.
Gottschewski's medium (Gottschewski, 1960)
The concentrations are given as mg/l,OOO ml medium.
L-Arginine 170, L-cysteine 40, L-cystine 3, glycine 12, L-histidine 196, L-Ieucine 200, Llysine 140, o-methionine 50, L-phenylalanine 200, L-threonine 120, L-tryptophan 20,
glucose 1,600, mannose 1,600, fructose 1,600, choline 5, folic acid 0.5, inositol 1.0, nicotinic acid 5.0, pantothenate 1.0, thiamine 0.5, riboflavin 0.5, pyridoxine 1.0, DPN 10,
TPN 5, adenine deoxyribonucleoside 0.5, guanine deoxyribonucleoside 0.5, uridine-5triphosphate O.5,deoxycytidine diphosphate 0.5,deoxy-5-methylcytidilic acid 0.5, thymidine diphosphate 0.5, ATP 5.0, RNA 50, DNA 50, NaCI 5,000, sodium acetate 50,
Na z HP0 4 ·5H z O 10,000, KC1300, CaCl z ·2H z O 20, MgCl z ·6H z O 300, MgS0 4 ·7H z O 300, glutathione 500, peptone 1,000, glycylglycine 1,000, casein hydrolysate 5,000, and yeast
extract 15. The pH of the medium is adjusted to 7.0-7.2 with 1 N NaOH and 1 N HCl.
Grace's medium (GMA; Grace, 1962)
The concentrations are given as mg/l,OOO ml medium.
NaH z P0 4 ·2HzO 1,140, NaHC0 3 350, KCI2,240, MgCl z ·6H z O 2,280, MgS0 4 ·7H z O 2,780,
CaCl z l,OOO, L-a-alanine 225, ~-alanine 200, L-arginine hydrochloride 700, L-asparagine
350, L-aspartic acid 350, L-cystine hydrochloride 25, L-glutamic acid 600, L-glutamine
600, glycine 650, L-histidine 2,500, L-isoleucine 50, L-Ieucine 75, L-Iysine hydrochloride
625, L-methionine 50, L-phenylalanine 150, L-proline 350, oL-serine 1,100, L-threonine
175, L-tryptophan 100, L-tyrosine 50, L-valine 100, sucrose 26,680, glucose 700, fructose
400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60, fumaric acid 55, thiamine
hydrochloride 0.02, riboflavin 0.02, calcium pantothenate 0.02, pyridoxine hydrochloride 0.02, p-aminobenzoic acid 0.02, folic acid 0.02, niacin 0.02, isoinositol 0.02, biotin
0.01, choline chloride 0.2, penicillin G sodium salt 30, and streptomycin sulfate 100,
made up to 1,000 ml with distilled water. The pH is adjusted to 6.5 with KOH. The
medium is used by supplementing it with 3% heat-inactivated hemolymph of Lepidoptera or 10% FBS.
407
200 ml, and dihydrostreptomycin sulfate 100, made up to 1,000 ml with distilled water.
The pH of the medium is adjusted to 6.2 with KOH.
Dulbecco's modified Eagle's medium (Morton, 1970)
The concentrations are given as mg/l,OOO ml medium.
CaClz 200, Fe(N03)3·9HzO 0.1, KCI 400, MgS04 97.67, NaCI 6,400, NaHC0 3 3,700,
NaH z P0 4 ·H z O 125, glucose 1,000, phenol red 15, sodium pyruvate 110, L-arginine hydrochloride 84, L-cystine dihydrochloride 62.57, L-glutamine 584, glycine 30, L-histidine hydrochloride monohydrate 42, L-isoleucine 104.8, L-Ieucine 104.8, L-Iysine hydrochloride 146.2, L-methionine 30, L-phenylalanine 66, L-serine 42, L-threonine 95.2,
tryptophan 16, L-tyrosine 2Na·2HzO 103.79, L-valine 93.6, o-calcium pantothenate 4.0,
choline chloride 4.0, folic acid 4.0, myoinositol 7.0, niacinamide 4.0, pyridoxal hydrochloride 4.0, riboflavin 0.4, and thiamine hydrochloride 4.0.
Durchon and Schaller's medium for Annelida (Durchon and Schaller, 1963)
Mix the following: 7 drops of 1 % agar in sterilized natural seawater containing 0.1 %
glucose, 3 drops of sterilized seawater containing 0.1% glucose, 1 drop of 50% egg
albumin in sterilized seawater containing 0.1 % glucose, 1 drop of horse serum (Difco
Laboratories), 1 drop of 50% chicken embryo extract in sterilized seawater containing
0.1 % glucose, 100 IV of penicillin G, and 0.009 mg of streptomycin.
Gottschewski's medium (Gottschewski, 1960)
The concentrations are given as mg/l,OOO ml medium.
L-Arginine 170, L-cysteine 40, L-cystine 3, glycine 12, L-histidine 196, L-Ieucine 200, Llysine 140, o-methionine 50, L-phenylalanine 200, L-threonine 120, L-tryptophan 20,
glucose 1,600, mannose 1,600, fructose 1,600, choline 5, folic acid 0.5, inositol 1.0, nicotinic acid 5.0, pantothenate 1.0, thiamine 0.5, riboflavin 0.5, pyridoxine 1.0, DPN 10,
TPN 5, adenine deoxyribonucleoside 0.5, guanine deoxyribonucleoside 0.5, uridine-5triphosphate O.5,deoxycytidine diphosphate 0.5,deoxy-5-methylcytidilic acid 0.5, thymidine diphosphate 0.5, ATP 5.0, RNA 50, DNA 50, NaCI 5,000, sodium acetate 50,
Na z HP0 4 ·5H z O 10,000, KC1300, CaCl z ·2H z O 20, MgCl z ·6H z O 300, MgS0 4 ·7H z O 300, glutathione 500, peptone 1,000, glycylglycine 1,000, casein hydrolysate 5,000, and yeast
extract 15. The pH of the medium is adjusted to 7.0-7.2 with 1 N NaOH and 1 N HCl.
Grace's medium (GMA; Grace, 1962)
The concentrations are given as mg/l,OOO ml medium.
NaH z P0 4 ·2HzO 1,140, NaHC0 3 350, KCI2,240, MgCl z ·6H z O 2,280, MgS0 4 ·7H z O 2,780,
CaCl z l,OOO, L-a-alanine 225, ~-alanine 200, L-arginine hydrochloride 700, L-asparagine
350, L-aspartic acid 350, L-cystine hydrochloride 25, L-glutamic acid 600, L-glutamine
600, glycine 650, L-histidine 2,500, L-isoleucine 50, L-Ieucine 75, L-Iysine hydrochloride
625, L-methionine 50, L-phenylalanine 150, L-proline 350, oL-serine 1,100, L-threonine
175, L-tryptophan 100, L-tyrosine 50, L-valine 100, sucrose 26,680, glucose 700, fructose
400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60, fumaric acid 55, thiamine
hydrochloride 0.02, riboflavin 0.02, calcium pantothenate 0.02, pyridoxine hydrochloride 0.02, p-aminobenzoic acid 0.02, folic acid 0.02, niacin 0.02, isoinositol 0.02, biotin
0.01, choline chloride 0.2, penicillin G sodium salt 30, and streptomycin sulfate 100,
made up to 1,000 ml with distilled water. The pH is adjusted to 6.5 with KOH. The
medium is used by supplementing it with 3% heat-inactivated hemolymph of Lepidoptera or 10% FBS.