Composition of Salt Solutions and Culture Media
409
440, and FBS 130 m!. The pH of the medium should be 7.0-7.2 and the osmolarity
155 mOsmollkg.
Haliotis rufescens culture medium (Naganuma et al., 1994)
Leibovitz's L-15 medium (Gibco) with the following components added: NaCI3% (w/
v) final concentration (originally 0.85%), L-glutamine 2 mM final concentration,
NaHC0 3 2 mM final concentration, antibiotic solution (100x; amphotericin B 25 mgt
ml, ampicillin 5 mg/ml, penicillin G 10,000 IU/ml, and streptomycin sulfate 10 mg/ml),
lipid mixture (1,OOOx; Sigma), yeast extract 200 mg/ml (100x; Sigma), calf serum (1020x). The final pH of the solution should be 7.6 and final osmolarity 1,080 mOsmollkg.
Heacox et al?s Medium G (Heacox et aI., 1983)
Natural seawater (sterile) 742 ml, distilled water 218 ml, NaCI 6,390 mg, NaHC0 3
1,000 mg, KCl340 mg, CaCl z ·2H z O 169 mg, glucose 1,000 mg, galactose 500 mg, fucose
(D) 250 mg, fucose (L) 250 mg, gentamicin 1.0 mg, penicillin 300,000 IU, Nereis coelomic fluid ultrafiltrate 0-20 ml, and Nereis vitellin 0 or 500 mg.
Honeybee medium (Kaatz et al., 1985)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 1,580, KCI 1,790, CaCl z ·2H z O 1,029, MgCl z ·6H z O 406, MgS0 4 ·7H z O 493,
NaH z P0 4 ·H z O 690, NaHC0 3 336, L-a-alanine 300, L-arginine 500, L-asparagine 200, Laspartic acid 100, L-cysteine 50, L-glutamic acid 250, L-glutamine 1,000, glycine 200, Lhistidine 200, L-isoleucine 100, L-Iysine 550, L-methionine 50, L-phenylalanine 100, Lproline 3,300, DL-serine 200, L-threonine 150, L-tryptophan 50, L-tyrosine 100, L-valine
150, glucose 4,000, fructose 2,500, sucrose 55,370, PIPES 7,560, bovine serum albumin
50,000, penicillin G ISO, streptomycin 250. The pH is adjusted to 6.70 with 1 N KOH
and 1 N HC!.
Hydra growth medium (Yu-Ying et aI., 1963)
Eagle's medium modified by Swin and Parker (1958) 10 m1,horse serum 100 ml,penicillin 500 IU, Fumidil B 200 mg, NaCI 5.8 mg, MgS0 4 ·7H z O 100 mg, NaHC0 3 168 mg,
KCI 7.5 mg, CaCl z III mg, dextrose 1,000 mg, phenol red 10 mg, streptomycin 12.5 mg,
triple-distilled water 800 ml, and 0.15 M phosphate buffer, to maintain the pH of the
medium at 6.8, made up to 1,000 ml with distilled water.
IPL-41 medium (Weiss et aI., 1981)
The concentrations are given as mg/l,OOO ml culture medium.
NaH z P0 4 ·HzO 1,160, NaHC03 350, KCl1,200, CaCl z 500, MgS0 4 ·7H z O 1,880, B-alanine
300, L-arginine hydrochloride 800, L-asparagine 1,300, L-aspartic acid 1,300, L-cystine
100, L-glutamic acid 1,500, L-glutamine 1,000, glycine 200, L-histidine 200, hydroXY-Lproline 800, L-isoleucine 750, L-leucine 250, L-Iysine hydrochloride 700, L-methionine
1,000, L-proline 500, L-phenylalanine 1,000, oL-serine 400, L-threonine 200, L-tryptophan
100, L-tyrosine 250, L-valine 500, sucrose 16,500, glucose 2,500, maltose 1,000, malic
acid 53.6, a-ketoglutaric acid 29.6, succinic acid 4.8, fumaric acid 4.4, thiamine hydrochloride 0.08, riboflavin 0.08, calcium pantothenate 0.008, pyridoxine hydrochloride
O.4,p-aminobenzoic acid 0.32, folic acid 0.08, niacin 0.16, inositol 0.4, biotin 0.16, cyanocobalamin 0.24, choline chloride 20, ZnClz 0.04, MnCl2·4H 2 0 0.02, CuCl 2 ·2H z O 0.2,
(NH4)Mo70Z4·4H20 0.04, CoClz·6HzO 0.05, and FeS04·7H20 0.55. The osmolarity of the
medium should be 360-375 mOsmollkg. FBS 10% may be added to the medium.
409
440, and FBS 130 m!. The pH of the medium should be 7.0-7.2 and the osmolarity
155 mOsmollkg.
Haliotis rufescens culture medium (Naganuma et al., 1994)
Leibovitz's L-15 medium (Gibco) with the following components added: NaCI3% (w/
v) final concentration (originally 0.85%), L-glutamine 2 mM final concentration,
NaHC0 3 2 mM final concentration, antibiotic solution (100x; amphotericin B 25 mgt
ml, ampicillin 5 mg/ml, penicillin G 10,000 IU/ml, and streptomycin sulfate 10 mg/ml),
lipid mixture (1,OOOx; Sigma), yeast extract 200 mg/ml (100x; Sigma), calf serum (1020x). The final pH of the solution should be 7.6 and final osmolarity 1,080 mOsmollkg.
Heacox et al?s Medium G (Heacox et aI., 1983)
Natural seawater (sterile) 742 ml, distilled water 218 ml, NaCI 6,390 mg, NaHC0 3
1,000 mg, KCl340 mg, CaCl z ·2H z O 169 mg, glucose 1,000 mg, galactose 500 mg, fucose
(D) 250 mg, fucose (L) 250 mg, gentamicin 1.0 mg, penicillin 300,000 IU, Nereis coelomic fluid ultrafiltrate 0-20 ml, and Nereis vitellin 0 or 500 mg.
Honeybee medium (Kaatz et al., 1985)
The concentrations are given as mg/l,OOO ml culture medium.
NaCI 1,580, KCI 1,790, CaCl z ·2H z O 1,029, MgCl z ·6H z O 406, MgS0 4 ·7H z O 493,
NaH z P0 4 ·H z O 690, NaHC0 3 336, L-a-alanine 300, L-arginine 500, L-asparagine 200, Laspartic acid 100, L-cysteine 50, L-glutamic acid 250, L-glutamine 1,000, glycine 200, Lhistidine 200, L-isoleucine 100, L-Iysine 550, L-methionine 50, L-phenylalanine 100, Lproline 3,300, DL-serine 200, L-threonine 150, L-tryptophan 50, L-tyrosine 100, L-valine
150, glucose 4,000, fructose 2,500, sucrose 55,370, PIPES 7,560, bovine serum albumin
50,000, penicillin G ISO, streptomycin 250. The pH is adjusted to 6.70 with 1 N KOH
and 1 N HC!.
Hydra growth medium (Yu-Ying et aI., 1963)
Eagle's medium modified by Swin and Parker (1958) 10 m1,horse serum 100 ml,penicillin 500 IU, Fumidil B 200 mg, NaCI 5.8 mg, MgS0 4 ·7H z O 100 mg, NaHC0 3 168 mg,
KCI 7.5 mg, CaCl z III mg, dextrose 1,000 mg, phenol red 10 mg, streptomycin 12.5 mg,
triple-distilled water 800 ml, and 0.15 M phosphate buffer, to maintain the pH of the
medium at 6.8, made up to 1,000 ml with distilled water.
IPL-41 medium (Weiss et aI., 1981)
The concentrations are given as mg/l,OOO ml culture medium.
NaH z P0 4 ·HzO 1,160, NaHC03 350, KCl1,200, CaCl z 500, MgS0 4 ·7H z O 1,880, B-alanine
300, L-arginine hydrochloride 800, L-asparagine 1,300, L-aspartic acid 1,300, L-cystine
100, L-glutamic acid 1,500, L-glutamine 1,000, glycine 200, L-histidine 200, hydroXY-Lproline 800, L-isoleucine 750, L-leucine 250, L-Iysine hydrochloride 700, L-methionine
1,000, L-proline 500, L-phenylalanine 1,000, oL-serine 400, L-threonine 200, L-tryptophan
100, L-tyrosine 250, L-valine 500, sucrose 16,500, glucose 2,500, maltose 1,000, malic
acid 53.6, a-ketoglutaric acid 29.6, succinic acid 4.8, fumaric acid 4.4, thiamine hydrochloride 0.08, riboflavin 0.08, calcium pantothenate 0.008, pyridoxine hydrochloride
O.4,p-aminobenzoic acid 0.32, folic acid 0.08, niacin 0.16, inositol 0.4, biotin 0.16, cyanocobalamin 0.24, choline chloride 20, ZnClz 0.04, MnCl2·4H 2 0 0.02, CuCl 2 ·2H z O 0.2,
(NH4)Mo70Z4·4H20 0.04, CoClz·6HzO 0.05, and FeS04·7H20 0.55. The osmolarity of the
medium should be 360-375 mOsmollkg. FBS 10% may be added to the medium.