Composition of Salt Solutions and Culture Media
411
Laverdure's medium (Laverdure, 1971)
Laverdure's medium is made by mixing 1 part of horse serum (Difco Laboratories;
dissolve the content of one ampoule in 5 ml bidistilled water), 1 part chicken embryo
(11 days old) extract (Difco Laboratories; dissolve the contents of one ampoule in 8 ml
Laverdure's physiological solution), 1 part Laverdure's physiological solution containing 2% peptone (Merck, Darmstadt, Germany), and 2 parts Laverdure's physiological
solution containing 1 % agar. Penicillin (20,000 IU) and streptomycin (0.5 mg) are added
to 20 ml medium. The pH of the medium is adjusted to 6.84-7.1 with 5.5% NaHC0 3 •
Leloup's medium 1 (Leloup, 1964)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI6,500, KC1250, MgS04·7H20 350, CaCl2 300, trehalose 800, glucose 700, malic acid
500, a-ketoglutaric acid 250, succinic acid 50, fumaric acid 50, yeast extract 1,000, and
lactalbumin hydrolysate 10,000. The pH of the medium is adjusted to 7.0-7.1 with 1 N
NaOH.
Limulus organ culture medium (Bayer and Barlow, 1978)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 16,100, KCI 276, CaCI 2 ·2H 2 0 1,184, MgCI 2 ·6H 2 0 1,423, MgS0 4 ·7H 2 0 5,669, medium 199 with Hanks salts (lOx; Gibco) 100 ml, horse serum 100 ml (Gibco), glucose
2,703, L-glutamine 292, retinol* (2 gil, Sigma) 10 ml, tocopherol* (200 mg/l, Sigma)
10 ml,penicillin-streptomycin (10,000 U/ml) 20 ml,HEPES (50 mM,Sigma) 10 ml,and
TES (N-tris (Hydroxymethyl) methyl-2-aminoethane-sulfonic acid; 2-( (2Hydroxy-l,
I-bis (hydroxymethyl)-ethyl) amino) ethansulfonic acid; 50 mM, Sigma) 10 ml, made
up to 1,000 ml with distilled water. The pH should be adjusted to 7.3.
* Add as an alcoholic solution.
Medium A-12 (Tripp et aI., 1966)
Mix 5% casamino acid 1.0 ml, 10% yeast extract 10 ml, 5% lactalbumin hydrolysate
10 ml, calf serum 10 mI, and BSSO (see Salt Solutions) 969 ml. The pH of the medium
should be 7.2-7.4.
Medium 1779 for Schistosoma (Weller and Wheeldon, 1982)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
CaCI 2 ·2H 2 0 400, KCll,OOO, MgS0 4 ·7H20 400, NaCI3,000, NaHC0 3 2,500, NaH 2 P0 4 10,
NaOH 200, HEPES 2,380, ZnS0 4 ·7H 2 0 0.022, FeS0 4 ·7H 2 0 0.01, MnS0 4 ·4H 2 0 0.004,
CuS0 4 0.0005, CoS0 4 ·7H20 0.00044, H 3 B0 3 0.00011, L-alanine 1,000, DL-cysteine hydrochloride 50, L-glutamine 150, glycine 250, L-histidine 1,000, hydroXY-L-proline 500,
L-Iysine hydrochloride 100, L-ornithine hydrochloride 10, L-serine 250, Eagle's BME
essential amino acids (Gibco) 1 ml, glutathione 10, L-histidyl-L-alanine 25, glucose
12,000, trehalose 500, adenine 2.5, cytosine 0.5, guanine hydrochloride 1.0, thiamine
0.5, uracil 1.0, xanthine sodium salt 1.0, orotic acid sodium salt 50.0, a-ketoglutaric
acid sodium salt 200, DL-carnitine hydrochloride 5.0, choline chloride 20.0, menadione
sodium bisulfite/albumin* 1 flg, vitamin B12 10.0, Eagle's BME vitamin block (Gibco)
5.0 ml, dexamethasone 392.5 ng, insulin 5.0, triiodo-L-thyronine 10 flg, Cohn's fraction IV 100, FBS 20.0 ml, gelatin 1,000, yeast extract/serum** 1.0 ml, phenol red 20,
serotonin creatinine sulfate 0.4, and cholesterol/albumin***.
411
Laverdure's medium (Laverdure, 1971)
Laverdure's medium is made by mixing 1 part of horse serum (Difco Laboratories;
dissolve the content of one ampoule in 5 ml bidistilled water), 1 part chicken embryo
(11 days old) extract (Difco Laboratories; dissolve the contents of one ampoule in 8 ml
Laverdure's physiological solution), 1 part Laverdure's physiological solution containing 2% peptone (Merck, Darmstadt, Germany), and 2 parts Laverdure's physiological
solution containing 1 % agar. Penicillin (20,000 IU) and streptomycin (0.5 mg) are added
to 20 ml medium. The pH of the medium is adjusted to 6.84-7.1 with 5.5% NaHC0 3 •
Leloup's medium 1 (Leloup, 1964)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI6,500, KC1250, MgS04·7H20 350, CaCl2 300, trehalose 800, glucose 700, malic acid
500, a-ketoglutaric acid 250, succinic acid 50, fumaric acid 50, yeast extract 1,000, and
lactalbumin hydrolysate 10,000. The pH of the medium is adjusted to 7.0-7.1 with 1 N
NaOH.
Limulus organ culture medium (Bayer and Barlow, 1978)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 16,100, KCI 276, CaCI 2 ·2H 2 0 1,184, MgCI 2 ·6H 2 0 1,423, MgS0 4 ·7H 2 0 5,669, medium 199 with Hanks salts (lOx; Gibco) 100 ml, horse serum 100 ml (Gibco), glucose
2,703, L-glutamine 292, retinol* (2 gil, Sigma) 10 ml, tocopherol* (200 mg/l, Sigma)
10 ml,penicillin-streptomycin (10,000 U/ml) 20 ml,HEPES (50 mM,Sigma) 10 ml,and
TES (N-tris (Hydroxymethyl) methyl-2-aminoethane-sulfonic acid; 2-( (2Hydroxy-l,
I-bis (hydroxymethyl)-ethyl) amino) ethansulfonic acid; 50 mM, Sigma) 10 ml, made
up to 1,000 ml with distilled water. The pH should be adjusted to 7.3.
* Add as an alcoholic solution.
Medium A-12 (Tripp et aI., 1966)
Mix 5% casamino acid 1.0 ml, 10% yeast extract 10 ml, 5% lactalbumin hydrolysate
10 ml, calf serum 10 mI, and BSSO (see Salt Solutions) 969 ml. The pH of the medium
should be 7.2-7.4.
Medium 1779 for Schistosoma (Weller and Wheeldon, 1982)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
CaCI 2 ·2H 2 0 400, KCll,OOO, MgS0 4 ·7H20 400, NaCI3,000, NaHC0 3 2,500, NaH 2 P0 4 10,
NaOH 200, HEPES 2,380, ZnS0 4 ·7H 2 0 0.022, FeS0 4 ·7H 2 0 0.01, MnS0 4 ·4H 2 0 0.004,
CuS0 4 0.0005, CoS0 4 ·7H20 0.00044, H 3 B0 3 0.00011, L-alanine 1,000, DL-cysteine hydrochloride 50, L-glutamine 150, glycine 250, L-histidine 1,000, hydroXY-L-proline 500,
L-Iysine hydrochloride 100, L-ornithine hydrochloride 10, L-serine 250, Eagle's BME
essential amino acids (Gibco) 1 ml, glutathione 10, L-histidyl-L-alanine 25, glucose
12,000, trehalose 500, adenine 2.5, cytosine 0.5, guanine hydrochloride 1.0, thiamine
0.5, uracil 1.0, xanthine sodium salt 1.0, orotic acid sodium salt 50.0, a-ketoglutaric
acid sodium salt 200, DL-carnitine hydrochloride 5.0, choline chloride 20.0, menadione
sodium bisulfite/albumin* 1 flg, vitamin B12 10.0, Eagle's BME vitamin block (Gibco)
5.0 ml, dexamethasone 392.5 ng, insulin 5.0, triiodo-L-thyronine 10 flg, Cohn's fraction IV 100, FBS 20.0 ml, gelatin 1,000, yeast extract/serum** 1.0 ml, phenol red 20,
serotonin creatinine sulfate 0.4, and cholesterol/albumin***.