408
Appendix
Haliotis discus culture medium (Naganuma et aI., 1996)
Leibovitz's L-15 medium (Gibco) with the following components added: NaCI 1.5%
(w/v) final concentration (originally 0.85%), NaHC0 3 2 mM final concentration, antibiotic-antimycotic solution (lOOx; Gibco; amphotericin B 25 mg/ml, ampicillin 5 mgt
ml, and streptomycin sulfate 10 mg/ml), lipid mixture (lOOx; Sigma), and FBS (lOx;
GIBCO). The final pH of the culture medium should be 7.8.
Hansen's M-182 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI2,800, KC1150, CaCl 2 530, MgS0 4 ·7H 2 0 450, Na 2 HP0 4 70, NaHC0 3 50, L-alanine
6.0, L-arginine 4.2, L-asparagine 2.0, L-aspartic acid 6.0, L-cysteine 6.0, L-cystine 2.4, Lglutamic acid 10.0, L-glutamine 20.0, glycine 6.0, L-histidine 4.0, L-isoleucine 5.2, Lleucine 5.2, L-Iysine 5.8, L-methionine 1.5, L-phenylalanine 3.5, L-proline 6.0, L-serine
12.0, L-threonine 4.8, L-tryptophan 0.8, L-tyrosine 3.6, L-valine 4.7, galactose 2,000, glucose 860, trehalose 860, biotin 0.038, calcium folinate 0.038, o-calcium pantothenate
0.075, cyanocobalamin 0.038, folic acid 0.075, N-acetylglucosamine 0.150, niacin 0.075,
niacinamide 0.075, pantethine 0.038, p-aminobenzoic acid 0.075, pyridoxal hydrochloride 0.038, pyridoxamine hydrochloride 0.038, pyridoxine hydrochloride 0.075, riboflavin 0.075, thiamine hydrochloride 0.075, thioctic acid 0.038, Bacto-peptone (Difco
Laboratories) 4,500, and FBS 130 ml. The pH of the medium should be 7.0-7.2 and the
osmolarity 185 mOsmollkg.
Hansen's M-260 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 1,792, KCI 96, CaCl 2 339, MgS0 4 ·7H 2 0 288, Na 2 HP0 4 45, NaHC0 3 32, L-alanine
3.8, L-arginine 2.7, L-asparagine 1.3, L-aspartic acid 3.8, L-cysteine 3.8, L-cystine 1.5, Lglutamic acid 6.4, L-glutamine 20.0, glycine 3.8, L-histidine 2.6, L-isoleucine 3.3, L-Ieucine 3.3, L-Iysine 3.7, L-methionine 1.0, L-phenylalanine 2.1, L-proline 3.8, L-serine 7.7,
L-threonine 3.1, L-tryptophan 0.5, L-tyrosine 2.3, L-valine 3.0, galactose 1,280, glucose
550, trehalose 550, biotin 0.024, calcium folinate 0.024, o-calcium pantothenate 0.048,
cyanocobalamin 0.024, folic acid 0.048, N-acetylglucosamine 0.096, niacin 0.048, niacinamide 0.048, pantethine 0.024, p-aminobenzoic acid 0.048, pyridoxal hydrochloride
0.024, pyridoxamine hydrochloride 0.024, pyridoxine hydrochloride 0.048, riboflavin
0.048, thiamine hydrochloride 0.048, thioctic acid 0.024, adenosine-3'(2')phosphoric
acid 0.347, cytidine-3'(2')phosphoric acid 0.323,guanosine-3'(2')phosphoric acid 0.363,
uridine-3'(2')phosphoric acid 0.324, thymine 0.126,lactalbumin hydrolysate (EdaminType S; Humco-Sheffield) 4,500, and FBS 130 ml. The pH of the medium should be
7.0-7.2 and the osmolarity 154 mOsmollkg.
Hansen's S-301 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl462 KC1352, CaCl2 132, MgS04·7H20 814, Na2HP04 154, KH2P04 99, NaHC0 3 88,
L-arginine 88, L-aspartic acid 88, ~-alanine 110, L-cysteine 13, L-cystine 22, L-glutamic
acid 176, L-glutamine 396, glycine 55, L-histidine 88, L-isoleucine 33, L-Ieucine 33, Llysine 363, L-methionine 176, L-phenylalanine 33, L-proline 374, L-serine 55, L-threonine 77, L-tryptophan 22, L-tyrosine 110, L-valine 66, galactose 1,300, glucose 440, trehalose 440, fumaric acid 22, a-ketoglutaric acid 44, malic acid 22, succinic acid 22,
lactalbumin hydrolysate (Edamin-Type S; Humco-Sheffield) 4,500, yeast hydrolysate
Appendix
Haliotis discus culture medium (Naganuma et aI., 1996)
Leibovitz's L-15 medium (Gibco) with the following components added: NaCI 1.5%
(w/v) final concentration (originally 0.85%), NaHC0 3 2 mM final concentration, antibiotic-antimycotic solution (lOOx; Gibco; amphotericin B 25 mg/ml, ampicillin 5 mgt
ml, and streptomycin sulfate 10 mg/ml), lipid mixture (lOOx; Sigma), and FBS (lOx;
GIBCO). The final pH of the culture medium should be 7.8.
Hansen's M-182 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI2,800, KC1150, CaCl 2 530, MgS0 4 ·7H 2 0 450, Na 2 HP0 4 70, NaHC0 3 50, L-alanine
6.0, L-arginine 4.2, L-asparagine 2.0, L-aspartic acid 6.0, L-cysteine 6.0, L-cystine 2.4, Lglutamic acid 10.0, L-glutamine 20.0, glycine 6.0, L-histidine 4.0, L-isoleucine 5.2, Lleucine 5.2, L-Iysine 5.8, L-methionine 1.5, L-phenylalanine 3.5, L-proline 6.0, L-serine
12.0, L-threonine 4.8, L-tryptophan 0.8, L-tyrosine 3.6, L-valine 4.7, galactose 2,000, glucose 860, trehalose 860, biotin 0.038, calcium folinate 0.038, o-calcium pantothenate
0.075, cyanocobalamin 0.038, folic acid 0.075, N-acetylglucosamine 0.150, niacin 0.075,
niacinamide 0.075, pantethine 0.038, p-aminobenzoic acid 0.075, pyridoxal hydrochloride 0.038, pyridoxamine hydrochloride 0.038, pyridoxine hydrochloride 0.075, riboflavin 0.075, thiamine hydrochloride 0.075, thioctic acid 0.038, Bacto-peptone (Difco
Laboratories) 4,500, and FBS 130 ml. The pH of the medium should be 7.0-7.2 and the
osmolarity 185 mOsmollkg.
Hansen's M-260 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 1,792, KCI 96, CaCl 2 339, MgS0 4 ·7H 2 0 288, Na 2 HP0 4 45, NaHC0 3 32, L-alanine
3.8, L-arginine 2.7, L-asparagine 1.3, L-aspartic acid 3.8, L-cysteine 3.8, L-cystine 1.5, Lglutamic acid 6.4, L-glutamine 20.0, glycine 3.8, L-histidine 2.6, L-isoleucine 3.3, L-Ieucine 3.3, L-Iysine 3.7, L-methionine 1.0, L-phenylalanine 2.1, L-proline 3.8, L-serine 7.7,
L-threonine 3.1, L-tryptophan 0.5, L-tyrosine 2.3, L-valine 3.0, galactose 1,280, glucose
550, trehalose 550, biotin 0.024, calcium folinate 0.024, o-calcium pantothenate 0.048,
cyanocobalamin 0.024, folic acid 0.048, N-acetylglucosamine 0.096, niacin 0.048, niacinamide 0.048, pantethine 0.024, p-aminobenzoic acid 0.048, pyridoxal hydrochloride
0.024, pyridoxamine hydrochloride 0.024, pyridoxine hydrochloride 0.048, riboflavin
0.048, thiamine hydrochloride 0.048, thioctic acid 0.024, adenosine-3'(2')phosphoric
acid 0.347, cytidine-3'(2')phosphoric acid 0.323,guanosine-3'(2')phosphoric acid 0.363,
uridine-3'(2')phosphoric acid 0.324, thymine 0.126,lactalbumin hydrolysate (EdaminType S; Humco-Sheffield) 4,500, and FBS 130 ml. The pH of the medium should be
7.0-7.2 and the osmolarity 154 mOsmollkg.
Hansen's S-301 medium (Hansen, 1976)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl462 KC1352, CaCl2 132, MgS04·7H20 814, Na2HP04 154, KH2P04 99, NaHC0 3 88,
L-arginine 88, L-aspartic acid 88, ~-alanine 110, L-cysteine 13, L-cystine 22, L-glutamic
acid 176, L-glutamine 396, glycine 55, L-histidine 88, L-isoleucine 33, L-Ieucine 33, Llysine 363, L-methionine 176, L-phenylalanine 33, L-proline 374, L-serine 55, L-threonine 77, L-tryptophan 22, L-tyrosine 110, L-valine 66, galactose 1,300, glucose 440, trehalose 440, fumaric acid 22, a-ketoglutaric acid 44, malic acid 22, succinic acid 22,
lactalbumin hydrolysate (Edamin-Type S; Humco-Sheffield) 4,500, yeast hydrolysate