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CELL CULTURE MEDIA AND REAGENTS
Institute for Fermentation, Osaka
HSRRB |
| The followings are cell culture media and reagents used in the IFO animal cell bank. These are listed only for reference, and you are not necessary to use the items from the same makers in your laboratory. | |
| Sera | |
| Calf serum | Flow (Cat. No. 29-131-54) |
| Fetal bovine (calf) serum (FBS) | BioWhittaker (Cat. No. 14-501F), (in Japan, available from Wako, Cat. No. 536-33905 B-W) |
| Horse serum | GIBCO BRL (Cat. No. 16050-122) |
| Heat-inactivated sera | Sera were heated at 56 C for 30 min. |
| Media | |
| minimal essential medium (MEM) | GIBCO BRL (Cat. No. 12000-022) |
| Dulbecco's modified Eagle's Medium (D.MEM) | Dulbecco's modified Eagle Medium "Nissui" (2), Nissui, Japan (Cat. No. 05919) |
| Dulbecco's modified Eagle's Medium (high glucose) | Dulbecco's modified Eagle's medium with 4.5 g/L glucose |
| Eagle's minimal essential medium (E.MEM) | Eagle's MEM "Nissui" (3), Nissui, Japan (Cat. No. 05902) |
| Eagle's basal medium with Earle's BSS | Basal medium Eagle with Earle's salts, Flow (Cat. No. 10-001-20) |
| Eagle's basal medium with Hanks' BSS | Basal medium Eagle with Hanks' salts, Flow (Cat. No. 10-031-22) |
| Ham's F10 medium | Flow (Cat. No. 10-401-20) |
| Ham's F12 medium | Ham's F12 medium "Nissui", Nissui, Japan (Cat. No. 05910) |
| Ham's F12K medium | Flow (Cat. No. 72-121-22) |
| HB101 medium | Irvine Scientific (in Japan, available from IS Japan) (Cat. No. T151-000) |
| Iscove's modified Dulbecco's medium | GIBCO BRL (Cat. No. 12200-085) |
| McCoy's 5A medium | Flow (Cat. No. 10-551-20) |
| MCDB 104 medium | MCDB 104GK medium, Kyokuto Seiyaku, Japan. (Now, this medium is not available from Kyokuto. MCDB 104 is available from BioFluid Inc. (in Japan, from Cosmo Bio), Cat. No. 117-500.) |
| MCDB 107 medium | Kyokuto Seiyaku, Japan (Cat. No. 25200-2). (Now, this medium is not available from Kyokuto.) |
| MCDB 153 medium | Kyokuto Seiyaku, Japan (Cat. No. 25400-6). (Now, this medium is not available from Kyokuto. MCDB153 is available from Sigma, Cat. No. M7403.) |
| Medium 199 | Nissui, Japan (Cat. No. 05909) |
| NCTC 109 medium | GIBCO BRL (Cat. No. 21340-039) |
| NCTC 135 medium | Flow (Cat. No. 10-911-22) |
| RITC 80-7 medium | Kyokuto Seiyaku, Japan (Cat. No. 26100-4). (Now, this medium is not available from Kyokuto.) |
| RPMI 1640 mdium | RPMI 1640 medium "Nissui" (2), Nissui, Japan (Cat. No. 05918) |
| Supplements for media | |
| Basic FGF | recombinant human basic fibroblast growth factor, Roche Molecular Biochemicals (Boehringer Mannheim) (Cat. No. 1-123-149) |
| Biotin | (+)-biotin, Wako, Japan (Cat. No. 023-08711) |
| Bovine pituitary extract | P-Neurext(r) , Upstate Biotechnology (Cat. No. 02-103) |
| Bovine serum albumin | albumin, bovine fraction V, Sigma (Cat. No. A 4503) |
| Bromodeoxyuridine | 5-bromo-2'-deoxyuridine, Wako, Japan (Cat. No. 029-04271) |
| Dexamethasone | Wako, Japan (Cat. No. 041-18861). |
| EGF | mouse epidermal growth factor, Takara Biomedicals, Japan (Cat. No. T001A) |
| Endothelial cell growth supplements | Collaborative Biomedical Products (in Japan, available from Nippon Becton Dickinson) (Cat. No. CBP40006) |
| Ethanolamine | monoethanolamine, Wako, Japan (Cat. No. 012-12455) |
| G5 supplements | 100 x G5 supplements, GIBCO BRL (Cat. No. 17503-012) |
| Glutamine | 200 mM L-glutamine (100 x ), Flow (Cat. No. 16-801-49) |
| HAT | 13.61 mg/L hypoxanthine, 0.18 mg/L aminopterin, 3.88 mg/L thymidine, HAT 50 x concentrate, Flow (Cat. No. 16-808-49) |
| Heparin | heparin, sodium salt, Wako, Japan (Cat. No. 085-00134) |
| HEPES | Dojindo Laboratories, Japan (Cat. No. 342-01375) |
| Hydrocortisone | Wako, Japan (Cat. No. 082-02481). |
| Insulin | recombinant human insulin, Wako, Japan (Cat. No. 096-03443) or insulin from bovine pancreas, Wako, Japan (Cat. No. 097-02971) |
| Interleukin-2 | recombinant human IL-2, Sionogi Seiyaku |
| Lactalbumin hydrolysate | Wako, USA (Cat. No. 549-00225) |
| Na-pyruvate | 100 mM sodium pyruvate for MEM (100 x concentrate), Flow (Cat. No. 16-820-49) |
| Na-selenite | sodium selenite, Wako, Japan (Cat. No. 196-10841) |
| NEAA | Non-essential amino acid for Eagle's minimal essential medium, MEM 100 x non-essential amino acids, Flow (Cat. No. 16-810-49) |
| Phosphoethanolamine | o-phosphorylethanolamine, Sigma (Cat. No. P 0503) |
| Transferrin | transferrin (holo), highly purified from bovine plasma, Ito Ham (available from Wako), Japan (Cat. No.300-01291) |
| Triiodothyronine | 3,3',5-triiodo-L-thyronine, Wako, Japan (Cat. No. 533-07341 FLU) |
| Enzymes and related reagents | |
| Crystalline trypsin | trypsin, cryst., Wako, Japan (Cat. No. 207-09891) |
| Dispase | dispase II, Godo Shusei (available from Sanko Jyunyaku), Japan or Roche Molecular Biochemicals (Boehringer Mannheim) (Cat. No. 1-276-921) |
| EDTA | ethylenediamine tetraacetic acid disodium salt (or tetrasodium salt), Dojindo Laboratories, Japan (Cat. No. 345-01861, 343-01883) |
| Pronase | Roche Molecular Biochemicals (Boehringer Mannheim) (Cat. No. 165-921) |
| Soybean trypsin inhibitor | trypsin inhibitor type I S, Sigma (Cat. No. T 6522) |
| Trypsin | 2.5% (10 x) trypsin 1:250, Flow (Cat. No. 16-893-49) or GIBCO BRL (Cat. No. 15090-046). These are 2.5% trypsin in normal concentration of saline. |
| Saline | |
| Ca, Mg-free PBS (phosphate buffered saline) | PBS tablets, Flow (Cat. No. 28-103-06) |
| Hanks' BSS (balanced salt solution) | Hanks' solution "Nissui" (1), Nissui, Japan (Cat. No. 05905) |
| Attachment factors | |
| Collagen type I | Cellmatrix-IP, Nitta Gelatin (available from Asahi Techno Glass (Iwaki)), Japan (Cat. No. CELLMATRIX-1P-100) |
| Fibronectin | fibronectin from bovine plasma, Ito Ham (available from Wako), Japan (Cat. No. 300-00571) |
| Laminin | laminin, mouse, Collaborative Biomedical Products (in Japan, available from Nippon Becton Dickinson) (Cat. No. CBP40232) |
| Poly-L-lysine | poly-L-lysine hydrochloride, Wako, Japan (Cat. No. 164-13421) |
| Reagents for freezing cells | |
| DMSO | dimethylsulfoxide, Wako, Japan (Cat. No. 048-21985) |
| Methylcellulose | methylcellulose 4000 cP, Wako, Japan (Cat. No. 136-02155). Methylcellulose solution can be neither autoclaved nor filtrated with membrane filter. For preparing the sterile solution, autoclave the methylcellulose powder with magnetic stirring bar in a glass bottle, add precociously sterilized base medium after the bottle cools down, and stir overnight. |
| Formulation of freezing media | |
| For serum-containing culture | 10% DMSO, 20% serum (final concentration) in base medium |
| For serum-free culture* | 10% DMSO, 0.1% methylcellulose in base medium(freezing at ca -1oC / min.) |
| * Ohno, T. et al. Cytotechnology, 1, 257 (1988). | |