Composition of Salt Solutions and Culture Media
413
chloride 1.0, riboflavin 0.1, inositol 1.8, vitamin B12 10.0, nicotinic acid 2.0, p-aminobenzoic acid 2.0, pyridoxine hydrochloride 2.0, uridine 2.0, deoxyadenosine 10.0,
deoxycytidine 10.0, deoxyguanosine 10, thymidine 10.0, cytosine 1.0, glucuronolactone
1.8, glutathione 10.0, phenol red 20.0, and glucose 100.0, made up to 1,000 ml with
distilled water. The pH of the medium should be adjusted to 7.2-7.4 with 1 N KOH and
1 N HCl.
Medium X for Drosophila imaginal discs (Davis and Shearn, 1977)
The concentrations are given as mg/1,000 ml culture medium.
MgS04·7H20 1,230, MgCI2·6H20 3,192, KH2P04 200, K 2 HP0 4 256, KC1446, KHC0 3 300,
potassium citrate 312, potassium fumarate 800, potassium a-ketoglutarate 870, potassium acetate 750, sodium citrate 3,122, sodium 1,4-piperazine diethanesulfonate (PIPES)
3,604, calcium succinate monohydrate 518, lactalbumin hydrolysate 9,000, L-proline
1,000, L-tryptophan 100, ~-alanine 50, L-cysteine 100, L-serine 250, y-aminobutyric acid
70, Bacto-peptone 5,000, trehalose 4,000, glucose 4,400, L-a-phosphatidylcholine,
dimyristoyl1.0, L-a-phosphatidylcholine, distearoyl1.0, cholesterol 2.6, TC-Yeastolate
2,000, vitamin A (trans-retinol) 0.1, vitamin E (o-a-tocopherol) 0.1, sodium a-glycerophosphate 1,620, calcium phosphorylcholine 1,290, phosphorylethanolamine 70,
BSA fraction V 10,000, polyvinylpyrrolidone (MW = 10,000) 250, polyvinylpyrrolidone
(MW = 40,000) 1,000, and polyvinylpyrrolidone (MW = 360,000) 9,000. The pH of the
medium should be adjusted to 6.8 with 1 N KOH and 1 N HCl.
This medium can be used following supplementation with Altosid (a juvenile hormone
analog; Zoecon) at a concentration of 31.0 ng/lOO ml medium and bovine insulin at
0.04 units/ 1 00 ml medium. Furthermore, 100 ml of this medium is conditioned either
by preincubation with the fat bodies of five larvae or by co culturing of the discs with
fat bodies.
MGM-434, MGM-443, MGM-448, MGM-450 and MGM-464
MGM-434 (chemically defined medium)
The concentrations are given as mg/1,OOO ml medium.
NaH2P04·2H 2 0 958, NaHC0 3 292,KCl1,875,CaCI 2 833, MgCI 2 ·6H 2 0 1,917, MgS0 4 ·7H 2 0
2,333, glucose 3,333, fructose 417, sucrose 22,080, malic acid, 558, a-ketoglutaric acid
308, succinic acid 50, fumaric acid 46, L-a-alanine 263, ~-alanine 167, L-arginine hydrochloride 583, L-asparagine 293, L-aspartic acid 293, L-cystine 2l, L-glutamic acid
500, L-glutamine 500, glycine 542, L-histidine 2,083, L-isoleucine 42, L-Ieucine 63, Llysine hydrochloride 521, L-methionine 42, L-phenylalanine 125, L-proline 292, oL-serine
917, L-threonine 146, L-tryptophan 83, L-tyrosine 42, L-valine 83, thiamine hydrochloride 0.16, riboflavin 0.16, pyridoxine hydrochloride 0.16, niacin 0.16, calcium pantothenate 0.16, biotin 0.08, folic acid 0.16, isoinositol 0.16,p-aminobenzoic acid 0.16, choline chloride 1.6, polyvinylpyrrolidone K-90 500. The pH of the medium should be
adjusted to 6.5 with concentrated KOH.
MGM-443 (Mitsuhashi, 1980)
This is MGM-434 medium modified by the addition of 10% FBS. The pH of the medium is adjusted to 6.3 with concentrated KOH solution.
MGM-448 (Mitsuhashi, 1984)
This is MGM-443 medium fortified by the addition of inosine 200 mg/l, cytochrome c
100 mg/I, fetuin 20 mg/l, and bovine plasma albumin fraction V 10,000 mg/l, FBS 100 mll
413
chloride 1.0, riboflavin 0.1, inositol 1.8, vitamin B12 10.0, nicotinic acid 2.0, p-aminobenzoic acid 2.0, pyridoxine hydrochloride 2.0, uridine 2.0, deoxyadenosine 10.0,
deoxycytidine 10.0, deoxyguanosine 10, thymidine 10.0, cytosine 1.0, glucuronolactone
1.8, glutathione 10.0, phenol red 20.0, and glucose 100.0, made up to 1,000 ml with
distilled water. The pH of the medium should be adjusted to 7.2-7.4 with 1 N KOH and
1 N HCl.
Medium X for Drosophila imaginal discs (Davis and Shearn, 1977)
The concentrations are given as mg/1,000 ml culture medium.
MgS04·7H20 1,230, MgCI2·6H20 3,192, KH2P04 200, K 2 HP0 4 256, KC1446, KHC0 3 300,
potassium citrate 312, potassium fumarate 800, potassium a-ketoglutarate 870, potassium acetate 750, sodium citrate 3,122, sodium 1,4-piperazine diethanesulfonate (PIPES)
3,604, calcium succinate monohydrate 518, lactalbumin hydrolysate 9,000, L-proline
1,000, L-tryptophan 100, ~-alanine 50, L-cysteine 100, L-serine 250, y-aminobutyric acid
70, Bacto-peptone 5,000, trehalose 4,000, glucose 4,400, L-a-phosphatidylcholine,
dimyristoyl1.0, L-a-phosphatidylcholine, distearoyl1.0, cholesterol 2.6, TC-Yeastolate
2,000, vitamin A (trans-retinol) 0.1, vitamin E (o-a-tocopherol) 0.1, sodium a-glycerophosphate 1,620, calcium phosphorylcholine 1,290, phosphorylethanolamine 70,
BSA fraction V 10,000, polyvinylpyrrolidone (MW = 10,000) 250, polyvinylpyrrolidone
(MW = 40,000) 1,000, and polyvinylpyrrolidone (MW = 360,000) 9,000. The pH of the
medium should be adjusted to 6.8 with 1 N KOH and 1 N HCl.
This medium can be used following supplementation with Altosid (a juvenile hormone
analog; Zoecon) at a concentration of 31.0 ng/lOO ml medium and bovine insulin at
0.04 units/ 1 00 ml medium. Furthermore, 100 ml of this medium is conditioned either
by preincubation with the fat bodies of five larvae or by co culturing of the discs with
fat bodies.
MGM-434, MGM-443, MGM-448, MGM-450 and MGM-464
MGM-434 (chemically defined medium)
The concentrations are given as mg/1,OOO ml medium.
NaH2P04·2H 2 0 958, NaHC0 3 292,KCl1,875,CaCI 2 833, MgCI 2 ·6H 2 0 1,917, MgS0 4 ·7H 2 0
2,333, glucose 3,333, fructose 417, sucrose 22,080, malic acid, 558, a-ketoglutaric acid
308, succinic acid 50, fumaric acid 46, L-a-alanine 263, ~-alanine 167, L-arginine hydrochloride 583, L-asparagine 293, L-aspartic acid 293, L-cystine 2l, L-glutamic acid
500, L-glutamine 500, glycine 542, L-histidine 2,083, L-isoleucine 42, L-Ieucine 63, Llysine hydrochloride 521, L-methionine 42, L-phenylalanine 125, L-proline 292, oL-serine
917, L-threonine 146, L-tryptophan 83, L-tyrosine 42, L-valine 83, thiamine hydrochloride 0.16, riboflavin 0.16, pyridoxine hydrochloride 0.16, niacin 0.16, calcium pantothenate 0.16, biotin 0.08, folic acid 0.16, isoinositol 0.16,p-aminobenzoic acid 0.16, choline chloride 1.6, polyvinylpyrrolidone K-90 500. The pH of the medium should be
adjusted to 6.5 with concentrated KOH.
MGM-443 (Mitsuhashi, 1980)
This is MGM-434 medium modified by the addition of 10% FBS. The pH of the medium is adjusted to 6.3 with concentrated KOH solution.
MGM-448 (Mitsuhashi, 1984)
This is MGM-443 medium fortified by the addition of inosine 200 mg/l, cytochrome c
100 mg/I, fetuin 20 mg/l, and bovine plasma albumin fraction V 10,000 mg/l, FBS 100 mll