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Nutrient Mixture F-12 Ham Formulation

Nutrient Mixture F12 Ham
Nutrient Mixture F12 Ham HEPES Modification
Nutrient Mixture F12 Ham Kaighn's Modification

Ham's nutrient mixtures were originally developed to cultivate Chinese hamster ovary (CHO) cells, mouse L-cells, and HeLa cells. These mixtures were formulated for use with or without serum supplementation like fetal bovine serum depending on the cell type cultured, and does not contain growth factors.

Ham's F-12 Nutrient Mixture, a modification on Ham’s F10, has been used to grow CHO cells, primary rat hepatocytes, and rat prostate epithelial cells. Ham's F-12 has several important modifications - Coon's modification, developed for culturing hybrid cells produced by viral fusion, and Kaighn's modification, which has increased concentrations of select amino acids, pyruvate, and modified salts. Kaighn's modification was designed to support the growth of differentiated rat and chicken cells and primary human hepatocytes. Coon’s modification consists of 2x the standard concentration of amino acids and pyruvate and includes ascorbic acid and adjusted salt concentrations. The Coon’s formula also contains 0.863 mg/L zinc sulfate, which may render it unsuitable for culturing mouse L-cells.

Nutrient Mixture F-12 Ham

Component	N4888 [1x] g/L	N6658 [1x] g/L	N6760 g/L
Inorganic Salts
Calcium Chloride	0.0333	0.0333	0.0333
Cupric Sulfate • 5H₂O	0.0000025	0.0000025	0.0000025
Ferrous Sulfate • 7H₂O	0.000834	0.000834	0.000834
Magnesium Chloride	0.0576	0.0576	0.0576
Potassium Chloride	0.224	0.224	0.224
Sodium Bicarbonate	1.176	1.176	—
Sodium Chloride	7.599	7.599	7.599
Sodium Phosphate Dibasic (anhydrous)	0.14204	0.14204	0.14204
Zinc Sulfate • 7H₂O	0.000863	0.000863	0.000863
Amino Acids
L-Alanine	0.009	0.009	0.009
L-Arginine • HCl	0.211	0.211	0.211
L-Asparagine • H₂O	0.01501	0.01501	0.01501
L-Aspartic Acid	0.0133	0.0133	0.0133
L-Cysteine • HCl • H₂O	0.035	0.035	0.035
L-Glutamic Acid	0.0147	0.0147	0.0147
L-Glutamine	—	0.146	0.146
Glycine	0.00751	0.00751	0.00751
L-Histidine • 3HCl • H₂O	0.02096	0.02096	0.02096
L-Isoleucine	0.00394	0.00394	0.00394
L-Leucine	0.0131	0.0131	0.0131
L-Lysine • HCl	0.0365	0.0365	0.0365
L-Methionine	0.00448	0.00448	0.00448
L-Phenylalanine	0.00496	0.00496	0.00496
L-Proline	0.0345	0.0345	0.345
L-Serine	0.0105	0.0105	0.0105
L-Threonine	0.0119	0.0119	0.0119
L-Tryptophan	0.00204	0.00204	0.00204
L-Tyrosine • 2Na • 2H₂O	0.00778	0.00778	0.00778
L-Valine	0.0117	0.0117	0.0117
Vitamins
D-Biotin	0.0000073	0.0000073	0.000073
Choline Chloride	0.01396	0.01396	0.01396
Folic Acid	0.00132	0.00132	0.00132
myo-Inositol	0.018	0.018	0.018
Niacinamide	0.000037	0.000037	0.000037
D-Pantothenic Acid (hemicalcium)	0.00048	0.00048	0.00048
Pyridoxine • HCl	0.000062	0.000062	0.000062
Riboflavin	0.000038	0.000038	0.000038
Thiamine • HCl	0.00034	0.00034	0.00034
Vitamin B₁₂	0.00136	0.00136	0.00136
Other
D-Glucose	1.802	1.802	1.802
Hypoxanthine	0.00408	0.00408	0.00408
Linoleic Acid	0.000084	0.000084	0.000084
Phenol Red • Na	0.0013	0.0013	0.0013
Putrescine • HCl	0.000161	0.000161	0.000161
Pyruvic Acid • Na	0.11	0.11	0.11
Thioctic Acid	0.00021	0.00021	0.00021
Thymidine	0.00073	0.00073	0.00073
Add
L-Glutamine	0.146	—	—
Sodium Bicarbonate	—	—	1.176

Nutrient Mixture F-12 Ham HEPES Modification

Component	N8641 [1x] g/L
Inorganic Salts
Calcium Chloride	0.0333
Cupric Sulfate • 5H₂O	0.0000025
Ferrous Sulfate • 7H₂O	0.000834
Magnesium Chloride	0.0576
Potassium Chloride	0.224
Sodium Bicarbonate	1.176
Sodium Chloride	7.1
Sodium Phosphate Dibasic (anhydrous)	0.14204
Zinc Sulfate • 7H₂O	0.000863
Amino Acids
L-Alanine	0.009
L-Arginine • HCl	0.211
L-Asparagine • H₂O	0.01501
L-Aspartic Acid	0.0133
L-Cysteine • HCl • H₂O	0.035
L-Glutamic Acid	0.0147
Glycine	0.00751
L-Histidine • 3HCl • H₂O	0.02096
L-Isoleucine	0.00394
L-Leucine	0.0131
L-Lysine • HCl	0.0365
L-Methionine	0.00448
L-Phenylalanine	0.00496
L-Proline	0.0345
L-Serine	0.0105
L-Threonine	0.0119
L-Tryptophan	0.00204
L-Tyrosine • 2Na • 2H₂O	0.00778
L-Valine	0.0117
Vitamins
D-Biotin	0.0000073
Choline Chloride	0.01396
Folic Acid	0.00132
myo-Inositol	0.018
Niacinamide	0.000037
D-Pantothenic Acid (hemicalcium)	0.000238
Pyridoxine • HCl	0.000062
Riboflavin	0.000038
Thiamine • HCl	0.00034
Vitamin B₁₂	0.00136
Other
D-Glucose	1.802
HEPES	5.958
Hypoxanthine	0.00408
Linoleic Acid	0.000084
Phenol Red • Na	0.0013
Putrescine • HCl	1.000161
Pyruvic Acid • Na	0.11
Thioctic Acid	0.00021
Thymidine	0.00073
Add
L-Glutamine	0.146

Nutrient Mixture F-12 Ham Kaighn's Modification

Component	N3520 [1x] g/L
Inorganic Salts
Calcium Chloride	0.10207
Cupric Sulfate • 5H₂O	0.0000025
Ferrous Sulfate • 7H₂O	0.000834
Magnesium Chloride	0.04961
Magnesium Sulfate	0.19264
Potassium Chloride	0.28329
Potassium Phosphate Monobasic	0.058523
Sodium Phosphate Dibasic	0.11502
Sodium Chloride	7.5972
Zinc Sulfate • 7H₂O	0.00014375
Amino Acids
L-Alanine	0.017818
L-Arginine	0.4214
L-Asparagine • H₂O	0.03002
L-Aspartic Acid	0.02662
L-Cysteine • HCl • H₂O	0.07024
L-Glutamic Acid	0.02942
L-Glutamine	0.2922
Glycine	0.015014
L-Histidine • HCl • H₂O	0.04192
L-Isoleucine	0.007872
L-Leucine	0.02624
L-Lysine • HCl	0.07304
L-Methionine	0.008952
L-Phenylalanine	0.009912
L-Proline	0.06906
L-Serine	0.02102
L-Threonine	0.02382
L-Tryptophan	0.004084
L-Tyrosine	0.010872
L-Valine	0.02342
Vitamins
D-Biotin	0.00007329
Choline Chloride	0.01396
Folic Acid	0.0013242
Hypoxanthine	0.004083
myo-Inositol	0.01802
Niacinamide	0.00003663
D-Pantothenic Acid (hemicalcium)	0.000477
Putrescine • 2HCl	0.0003222
Pyridoxine • HCl	0.00006168
Riboflavin	0.00003764
Thiamine • HCl	0.0003373
Thymidine	0.0007266
Vitamin B₁₂	0.0013554
Other
D+-Glucose	1.26
Phenol Red • Na	0.00331806
Pyruvic Acid • Na	0.22
DL-6,8-Thioctic Acid	0.0002063
Add
Sodium Bicarbonate	2.5