414
Appendix
1, or Antheraea pernyi hemolymph 30 mlll. The pH of the medium is adjusted to 6.3
with concentrated KOH solution.
MGM-450 (Mitsuhashi and Inoue, 1988)
This medium is prepared by adding 3,000 mg tryptose phosphate broth to 1,000 ml of
MGM-448 medium. The pH is adjusted to 6.3 with concentrated KOH solution.
This medium can also be used by adding FBS at 10%, or FBS 5% and Antheraea pernyi
hemolymph 3%.
MGM-464 (Mitsuhashi, 2001)
This medium is prepared by adding 3,000 mg tryptose phosphate broth, 3,000 mg TCYeastolate (Difco), and 3,000 mg lactalbumin hydrolysate to 1,000 ml of MGM-448
medium. The pH of the medium is adjusted to 6.3 with concentrated KOH solution.
This medium is used by adding FBS at 20%.
MM medium (Mitsuhashi and Maramorosch, 1964)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl 7,000, KCl 200, CaC12·2H20 200, MgC12·6H20 100, NaH 2 P0 4 200, NaHC0 3 120,
glucose 4,000, lactalbumin hydrolysate 6,500, TC-Yeastolate 5,000, and FBS 0-200 ml,
made up to 1,000 ml with distilled water. The pH is adjusted to 6.3 with KOH (the pH
of the medium is already around 6.3 without any adjustment). The medium can be
sterilized before the addition of FBS by autoclaving at 121°C for 15 min. The medium
can be used as a serum-free medium (= MM-SF medium).
Nudibranch culture medium (Tamse et aI., 1995)
This is Leibovitz's L-15 medium supplemented with 444 mM NaCl , 10.0 mM KCl,
28.0 mM MgC1 2 ·6H 2 0, 27.0 mM MgS0 4 ·7H 2 0, 2.0 mM NaHC0 3 , 35.0 mM D-glucose,
15.0 mM HEPES, 11.0 mM CaC1 2 ·2H 2 0, penicillin 50 lUI ml, and streptomycin 50 Ilg1
ml. The pH of the medium should be 7.4-7.5.
Ogura's medium (Ogura, 1989)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl 7,900, KCl 290, CaCl z ·2H 2 0 200, MgS0 4 ·7H 2 0 150, Na 2 HP0 4 ·12Hp 380, KH 2 P0 4
80, NaHC0 3 1,200, glucose 1,500, lactalbumin hydrolysate 500, TC-Yeastolate 250,
chicken embryo extract 2.5 ml, nematode hemolymph * 5 ml, penicillin 10 5 IV, and streptomycin 100, made up to 1,000 ml with distilled water.
*Nematode hemolymph is prepared from the female adults of Toxocara canis.
Oncomelania culture medium (Iwanaga et aI., 1985)
Vnless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
NaC12,800, KCl150, Na 2 HP0 4 70, MgS0 4 ·7H 2 0 450, CaC1 2 ·2H 2 0 530, NaHC0 3 50, galactose 5,000, glucose 5,000, trehalose 1,000, a-ketoglutaric acid 1,200, fumaric acid
800, malic acid 800, succinic acid 800, L-lysine hydrochloride 208, L-histidine 100, Larginine hydrochloride 144, L-aspartic acid 532, L-threonine 288, L-serine 360, L-glutamic
acid 516, L-proline 248, glycine 388, L-alanine 352, L-valine 104, L-methionine 88, Lisoleucine 116, L-leucine 292, L-tyrosine 128, L-phenylalanine 156, Eagle's MBE vitamin
(xl00; Gibco) 40 ml, and phenol red 5, made up to 1,000 ml with distilled water. Vse
the medium by adding heat-treated horse serum at a concentration of 10%. The pH of
the medium should be 7.0-7.2.
Appendix
1, or Antheraea pernyi hemolymph 30 mlll. The pH of the medium is adjusted to 6.3
with concentrated KOH solution.
MGM-450 (Mitsuhashi and Inoue, 1988)
This medium is prepared by adding 3,000 mg tryptose phosphate broth to 1,000 ml of
MGM-448 medium. The pH is adjusted to 6.3 with concentrated KOH solution.
This medium can also be used by adding FBS at 10%, or FBS 5% and Antheraea pernyi
hemolymph 3%.
MGM-464 (Mitsuhashi, 2001)
This medium is prepared by adding 3,000 mg tryptose phosphate broth, 3,000 mg TCYeastolate (Difco), and 3,000 mg lactalbumin hydrolysate to 1,000 ml of MGM-448
medium. The pH of the medium is adjusted to 6.3 with concentrated KOH solution.
This medium is used by adding FBS at 20%.
MM medium (Mitsuhashi and Maramorosch, 1964)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl 7,000, KCl 200, CaC12·2H20 200, MgC12·6H20 100, NaH 2 P0 4 200, NaHC0 3 120,
glucose 4,000, lactalbumin hydrolysate 6,500, TC-Yeastolate 5,000, and FBS 0-200 ml,
made up to 1,000 ml with distilled water. The pH is adjusted to 6.3 with KOH (the pH
of the medium is already around 6.3 without any adjustment). The medium can be
sterilized before the addition of FBS by autoclaving at 121°C for 15 min. The medium
can be used as a serum-free medium (= MM-SF medium).
Nudibranch culture medium (Tamse et aI., 1995)
This is Leibovitz's L-15 medium supplemented with 444 mM NaCl , 10.0 mM KCl,
28.0 mM MgC1 2 ·6H 2 0, 27.0 mM MgS0 4 ·7H 2 0, 2.0 mM NaHC0 3 , 35.0 mM D-glucose,
15.0 mM HEPES, 11.0 mM CaC1 2 ·2H 2 0, penicillin 50 lUI ml, and streptomycin 50 Ilg1
ml. The pH of the medium should be 7.4-7.5.
Ogura's medium (Ogura, 1989)
The concentrations are given as mg/l,OOO ml culture medium.
NaCl 7,900, KCl 290, CaCl z ·2H 2 0 200, MgS0 4 ·7H 2 0 150, Na 2 HP0 4 ·12Hp 380, KH 2 P0 4
80, NaHC0 3 1,200, glucose 1,500, lactalbumin hydrolysate 500, TC-Yeastolate 250,
chicken embryo extract 2.5 ml, nematode hemolymph * 5 ml, penicillin 10 5 IV, and streptomycin 100, made up to 1,000 ml with distilled water.
*Nematode hemolymph is prepared from the female adults of Toxocara canis.
Oncomelania culture medium (Iwanaga et aI., 1985)
Vnless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
NaC12,800, KCl150, Na 2 HP0 4 70, MgS0 4 ·7H 2 0 450, CaC1 2 ·2H 2 0 530, NaHC0 3 50, galactose 5,000, glucose 5,000, trehalose 1,000, a-ketoglutaric acid 1,200, fumaric acid
800, malic acid 800, succinic acid 800, L-lysine hydrochloride 208, L-histidine 100, Larginine hydrochloride 144, L-aspartic acid 532, L-threonine 288, L-serine 360, L-glutamic
acid 516, L-proline 248, glycine 388, L-alanine 352, L-valine 104, L-methionine 88, Lisoleucine 116, L-leucine 292, L-tyrosine 128, L-phenylalanine 156, Eagle's MBE vitamin
(xl00; Gibco) 40 ml, and phenol red 5, made up to 1,000 ml with distilled water. Vse
the medium by adding heat-treated horse serum at a concentration of 10%. The pH of
the medium should be 7.0-7.2.