US5232848A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Basal nutrient medium for cell culture
Assignee: Amicon Corp
•
Filing: 1989-11-16
•
Granted: 1993-08-03
•
Expires: 2009-11-16
Abstract
Abstract This defined basal nutrient medium is very effective in both high and low density culture of a wide variety of cell lines and cell types. The medium may be used for serum-free culture or supplemented with low levels of serum. The medium contains a buffer system formulated for air equilibration.
Inventors
- 👤 Richard A. Wolfe
- 👤 Aaron H. Heifetz
- 👤 Linda M. Custer
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 13:01
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0018; C12N5/0037; C12N2500/00; C12N2500/05; C12N2500/10; C12N2500/20; C12N2500/24; C12N2500/25
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
375 words
An air-equilibrated basal nutrient medium for in vitro mammalian cell culture, consisting essentially of the following: ______________________________________
D-Glucose 2 × 10.sup.-2 M
Na Pyruvate 1 × 10.sup.-3 M
CaCl.sub.2.2H.sub.2 O
1 × 10.sup.-3 M
CuSO.sub.4.5H.sub.2 O
3 × 10.sup.-9 M
FeSO.sub.4.7H.sub.2 O
1 × 10.sup.-6 M
Fe(NO.sub.3).sub.3.9H.sub.2 O
2 × 10.sup.-7 M
KCl 4 × 10.sup.-3 M
MgSO.sub.4.7H.sub.2 O
8 × 10.sup.-4 M
NaCl 1.05 × 10.sup.-1 M
Na.sub.2 HPO.sub.4.7H.sub.2 O
3 × 10.sup.-4 M
NaH.sub.2 PO.sub.4.2H.sub.2 O
6 × 10.sup.-4 M
Na.sub.2 SeO.sub.3.5H.sub.2 O
3 × 10.sup.-8 M
Na.sub.2 SiO.sub.3.9H.sub.2 O
1 × 10.sup.-5 M
(NH.sub.4)6Mo.sub.7 O.sub.24.4H.sub.2 O
3 × 10.sup.-9 M
NH.sub.4 VO.sub.3
5 × 10.sup.-10 M
NiSO.sub.4.6H.sub.2 O
3 × 10.sup.-10 M
ZnSO.sub.4.7H.sub.2 O
8 × 10.sup.-7 M
L-Arg 8 × 10.sup.-4 M
L-Cys HCl.H.sub.2 O
3 × 10.sup.-4 M
L-Gln 5 × 10.sup.-5 M
L-His HCl.H.sub.2 O
2 × 10.sup.-4 M
L-Hydroxy-Pro 1 × 10.sup.-4 M
L-Ile 6 × 10.sup.-4 M
L-Leu 6 × 10.sup.-4 M
L-Lys HCl 8 × 10.sup.-4 M
L-Met 1 × 10.sup.-3 M
L-Phe 3 × 10.sup.-4 M
L-Thr 6 × 10.sup.-4 M
L-Trp 6 × 10.sup.-5 M
L-Tyr (diNa.sup.+)2H.sub.2 O
3 × 10.sup.-4 M
L-Val 6 × 10.sup.-4 M
L-Ala 2 × 10.sup.-5 M
L-Asn.H.sub.2 O 3 × 10.sup.-4 M
L-Asp 2 × 10.sup.-5 M
L-Glu 2 × 10.sup.-5 M
Gly 3 × 10.sup.-5 M
L-Pro 2 × 10.sup.-4 M
L-Ser 3 × 10.sup.-4 M
Glutathione 1 × 10.sup.-6 M
Putrescine 2HCl 3 × 10.sup.-7 M
Biotin 3 × 10.sup.-8 M
D-Ca pantothenate
2 × 10.sup.-5 M
Folic acid 6 × 10.sup.-6 M
Folinic acid (Ca.sup.+).5H.sub.2 O
1 × 10.sup.-6 M
Niacinamide 3 × 10.sup.-5 M
(Nicotinamide)
p-Aminobenzoic acid
3 × 10.sup.-6 M
Pyridoxal HCl 1 × 10.sup.-5 M
Pyridoxine HCl 3 × 10.sup.-7 M
Riboflavin 8 × 10.sup.-7 M
Thiamine HCl 9 × 10.sup.-6 M
Vitamin B12 3 × 10.sup.-7 M
Adenine 1 × 10.sup.-6 M
Hypoxanthine (Na.sup.+)
7 × 10.sup.-6 M
Thymidine HCl 1 × 10.sup.-5 M
Choline chloride 1 × 10.sup.-4 M
Ethanolamine HCl 2 × 10.sup.-5 M
i-Inositol 1 × 10.sup.-4 M
Linoleic acid 1 × 10.sup.-7 M
Lipoic acid 2 × 10.sup.-7 M
Zwitterionic Buffer
2.5 × 10.sup.-2 M
NaOH 1.23 × 10.sup. -2 M
NaHCO.sub.3 3 × 10.sup.-3 M
______________________________________
Claim 2
Dependent Claim
375 words
An air-equilibrated basal nutrient medium for in vitro mammalian cell culture, consisting essentially of the following: ______________________________________
D-Glucose 2 × 10.sup.-2 M
Na Pyruvate 1 × 10.sup.-3 M
CaCl.sub.2.2H.sub.2 O
1 × 10.sup.-3 M
CuSO.sub.4.5H.sub.2 O
3 × 10.sup.-9 M
FeSO.sub.4.7H.sub.2 O
1 × 10.sup.-6 M
Fe(NO.sub.3).sub.3.9H.sub.2 O
2 × 10.sup.-7 M
KCl 4 × 10.sup.-3 M
MgSO.sub.4.7H.sub.2 O
8 × 10.sup.-4 M
NaCl 1.05 × 10.sup.-1 M
Na.sub.2 HPO.sub.4.7H.sub.2 O
3 × 10.sup.-4 M
NaH.sub.2 PO.sub.4.2H.sub.2 O
6 × 10.sup.-4 M
Na.sub.2 SeO.sub.3.5H.sub.2 O
3 × 10.sup.-8 M
Na.sub.2 SiO.sub.3.9H.sub.2 O
1 × 10.sup.-5 M
(NH.sub.4)6Mo.sub.7 O.sub.24.4H.sub.2 O
3 × 10.sup.-9 M
NH.sub.4 VO.sub.3
5 × 10.sup.-10 M
NiSO.sub.4.6H.sub.2 O
3 × 10.sup.-10 M
ZnSO.sub.4.7H.sub.2 O
8 × 10.sup.-7 M
L-Arg 8 × 10.sup.-4 M
L-Cys HCl.H.sub.2 O
3 × 10.sup.-4 M
L-Gln 5 × 10.sup.-5 M
L-His HCl.H.sub.2 O
2 × 10.sup.-4 M
L-Hydroxy-Pro 1 × 10.sup.-4 M
L-Ile 6 × 10.sup.-4 M
L-Leu 6 × 10.sup.-4 M
L-Lys HCl 8 × 10.sup.-4 M
L-Met 1 × 10.sup.-3 M
L-Phe 3 × 10.sup.-4 M
L-Thr 6 × 10.sup.-4 M
L-Trp 6 × 10.sup.-5 M
L-Tyr (diNa.sup.+)2H.sub.2 O
3 × 10.sup.-4 M
L-Val 6 × 10.sup.-4 M
L-Ala 2 × 10.sup.-5 M
L-Asn.H.sub.2 O 3 × 10.sup.-4 M
L-Asp 2 × 10.sup.-5 M
L-Glu 2 × 10.sup.-5 M
Gly 3 × 10.sup.-5 M
L-Pro 2 × 10.sup.-4 M
L-Ser 3 × 10.sup.-4 M
Glutathione 1 × 10.sup.-6 M
Putrescine 2HCl 3 × 10.sup.-7 M
Biotin 3 × 10.sup.-8 M
D-Ca pantothenate
2 × 10.sup.-5 M
Folic acid 6 × 10.sup.-6 M
Folinic acid (Ca.sup.+).5H.sub.2 O
1 × 10.sup.-6 M
Niacinamide 3 × 10.sup.-5 M
(Nicotinamide)
p-Aminobenzoic acid
3 × 10.sup.-6 M
Pyridoxal HCl 1 × 10.sup.-5 M
Pyridoxine HCl 3 × 10.sup.-7 M
Riboflavin 8 × 10.sup.-7 M
Thiamine HCl 9 × 10.sup.-6 M
Vitamin B12 3 × 10.sup.-7 M
Adenine 1 × 10.sup.-6 M
Hypoxanthine (Na.sup.+)
7 × 10.sup.-6 M
Thymidine HCl 1 × 10.sup.-5 M
Choline chloride 1 × 10.sup.-4 M
Ethanolamine HCl 2 × 10.sup.-5 M
i-Inositol 1 × 10.sup.-4 M
Linoleic acid 1 × 10.sup.-7 M
Lipoic acid 2 × 10.sup.-7 M
Zwitterionic Buffer
2.5 × 10.sup.-2 M
NaOH 1.23 × 10.sup. -2 M
NaHCO.sub.3 3 × 10.sup.-3 M
______________________________________
Claim 3
Dependent Claim
25 words
The nutrient medium of claim 1 wherein the zwitterionic buffer is selected from the group consisting of HEPES, MOPSO, BES, TAPSO and a combination thereof.
Claim 4
Dependent Claim
21 words
The nutrient medium of claim 1 which is supplemented with an inorganic iron source, and defined proteins, or a combination thereof.
Claim 5
Dependent Claim
28 words
The nutrient medium of claim 3 wherein said inorganic iron source is selected from the group consisting of ferric sulfate, ferric citrate, ferrous fumarate and ferrous ammonium sulfate.
Claim 6
Dependent Claim
22 words
The nutrient medium of claim 3 wherein said defined proteins are selected from the group consisting of albumin, insulin and iron-saturated transferrin.
Claim 7
Dependent Claim
17 words
The nutrient medium of claim 1 which has an osmolarity of about 285 to about 315 milli-osmols.
Claim 8
Dependent Claim
17 words
The nutrient medium of claim 6 which has an osmolarity of about 295 to about 305 milli-osmols.
Claim 9
Dependent Claim
16 words
The nutrient medium of claim 1 which has a pH of about 7.0 to about 8.0.
Claim 10
Dependent Claim
13 words
The nutrient medium of claim 8 which has a pH of about 7.2.
Claim 11
Dependent Claim
16 words
The nutrient medium of claim 8 which has a pH of about 7.35 to about 7.8.
Claim 12
Dependent Claim
17 words
The nutrient medium of claim 11 which has a sodium-to-potassium ratio of about 25 to about 35.
Claim 13
Dependent Claim
13 words
The nutrient medium of claim 11 in which said ratio is about 30.
Claim 14
Dependent Claim
18 words
The nutrient medium of claim 1 wherein the nutrient medium is supplemented with serum or another biological extract.
Claim 15
Dependent Claim
23 words
The nutrient medium of claim 13 in which said serum or extract is less than about one percent of the medium by volume.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-08-02 | 1997-02-01 | 1997-08-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-08-02 | 2001-02-01 | 2001-08-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-08-02 | 2005-02-01 | 2005-08-03 | $7700.00 | Window Closed / Lapsed |