Composition for maintaining organ and cell viability
The present invention is directed to methods of preserving mammalian heart, ex vivo. Nanoparticle compositions for carrying out the invention are also disclosed.
1. A method of preserving a mammalian heart, ex vivo, comprising contacting or perfusing a mammalian heart with an effective amount of an aqueous composition that comprises:
a first phase comprising a base nutritive medium; and
a second phase comprising nanoparticles that comprise an outer lipophilic coating and an inner hydrophilic core, wherein
a) the first phase comprises physiologically compatible concentrations of water soluble or dispersible nutrients, and physiological salts;
b) the nanoparticles of the second phase comprise lipids, fatty acids, sterols and free fatty acids; and
c) the two-phase composition has an osmolality of at least about 300 mOsM/kg, and wherein the nanoparticles have a mean diameter ranging from about 100 nm to about 300 nm, and wherein the hydrophilic inner core of the nanoparticle contains a solution or suspension comprising a moiety capable of binding and releasing oxygen, and the first phase comprises a composition of the following table:
g/Liter
RANGE
Adenosine
0.950-1.050
Adenosine 5′ Monophosphate
0.0019-0.0021
Adenosine Triphosphate
0.0019-0.0021
Allopurinol
0.133-0.147
B′ Nicotinamide Adenine Dinucleotide
0.038-0.042
Phosphate
B′Nicotinamide Adenine Dinucleotide
0.0019-0.0021
Calcium Chloride
0.152-0.168
Choline Chloride
0.0085-0.0094
Chondrotin Sulfate
0.0038-0.0042
Cocarboxylase
0.038-0.042
Coenzyme A
0.0095-0.00105
Cyclodextrin
0.475-0.525
Deoxyadenosine
0.038-0.042
Deoxycytidine
0.038-0.042
Deoxyguanosine
0.038-0.042
Dextran 70
33.25-36.75
Flavin Adenine Dinucleotide
0.038-0.042
Folic Acid
0.0026-0.0028
Glucose
3.800-4.200
Glutathione
0.950-1.050
Glycine
0.0179-0.0197.
2. The method of claim 1 , wherein the composition has an osmolality ranging from about 385-425 mOsM/kg.
3. The method of claim 1 , wherein the pH of the composition is from about 7.2 to about 7.4.
4. The method of claim 1 , wherein the nanoparticles of the composition have a mean diameter ranging from about 100 nm to about 300 nm.
5. The method of claim 4 , wherein the nanoparticles have a mean diameter ranging from about 100 nm to about 200 nm.
6. The method of claim 1 , wherein the composition further comprises cellular growth factors.
7. The method of claim 6 , wherein said cellular growth factors are selected from the group consisting of epithelial and endothelial growth factors, vascular endothelial growth factors, platelet derived endothelial growth factors, epithelial growth factors, hepatocyte growth factors, and mixtures thereof.
8. The method of claim 1 , wherein the composition comprises about equal amounts of the first phase and the second phase.
9. The method of claim 1 , wherein the outer lipophilic coating of the composition comprises a free fatty acid selected from the group consisting of oleic acid and linoleic acid, and combinations thereof.
10. The method of claim 1 , wherein the composition further comprises:
g/Liter
RANGE
Adenine HCl
0.00019-0.00021
vitamin B-12
0.00065-0.0007
Biotin
0.00000038-0.00000042
Cupric Sulfate
0.00000124-0.00000137
Ferric Nitrate
0.000048-0.000053
Ferric Sulfate
0.00048-0.00053
Putrescine HCl
0.000077-0.000085
Pyridoxine HCl
0.000029-0.000033
Riboflavin
0.00021-0.000231
Thymidine
0.00035-0.00039
Zinc Sulfate
0.00041-0.000454.
11. The method of claim 1 , wherein the composition further comprises:
g/Liter
RANGE
Heparin
0.171-0.189
HEPES
3.396-3.753
Hypoxanthine
0.002-0.0022
Inositol
0.0124-0.0137
Insulin
0.0095-0.0105
L-Alanine
0.00428-0.00473
L-Arginine
0.141-0.155
L-Asparagine
0.0076-0.0084
L-Aspartic Acid
0.064-0.070
L-Cysteine
0.0297-0.0329
L-Cystine
0.0167-0.0185
L-Glutamic Acid
0.007-0.0078
L-Glutamine
4.750-5.250
L-Histidine
0.030-0.033
L-Isoleucine
0.052-0.0572
L-Leucine
0.057-0.063
L-Lysine
0.0095-0.0105
L-Methionine
0.019-0.021
L-Phenylalanine
0.0337-0.0373
L-Proline
0.0164-0.0182
L-Serine
0.025-0.0276
L-Threonine
0.051-0.056.
12. The method of claim 1 , wherein the composition further comprises:
g/Liter
RANGE
L-Tryptophan
0.009-0.0095
L-Tyrosine
0.053-0.059
L-Valine
0.050-0.055
Magnesium Chloride
0.058-0.0643
Magnesium Sulfate
0.0475-0.0525
Mannose
3.135-3.465
Niacinamide
0.0019-0.0021
Pantothenic Acid
0.0021-0.0024
Potassium Chloride
0.296-0.328
Pyridoxal HCl
0.0019-0.0021
Pyruvic Acid
0.209-0.231
Sodium Bicarbonate
1.140-1.260
Sodium Chloride
6.650-7.350
Sodium Phosphate Dibasic
0.0676-0.0748
Sodium Phosphate Monobasic
0.0516-0.0570
Thiamine
0.0021-0.0023
Transferrin
0.00475-0.00525
Uridine
0.038-0.042
Uridine Triphosphate
0.038-0.042
Yeastolate Ultra-Filtered
38-42 ML.
13. The method of claim 1 , wherein the composition further comprises:
g/Liter
RANGE
L-Cystine
0.0167-0.0185
L-Tyrosine
0.053-0.059.
14. The method of claim 1 , wherein the second phase of the composition further comprises:
g/Liter
RANGE
Cholesterol
0.00475-0.00525
Cod Liver Oil
0.00095-0.00105
Epithelial Growth Factor
0.00000285-0.00000315
Hepatocyte Growth Factor
0.0000048-0.0000053
Hydrocortisone
0.00095-0.00105
Linoleic Acid
0.00095-0.00105
Linolenic Acid
0.00095-0.00105
Oleic Acid
0.00095-0.00105
Phosphatidylcholine
0.6%
Platelet Derived Endothelial Growth
0.00000095-0.00000105
Factor
Pluronic F-68
0.950-1.050
Prostaglandin E1
0.000042-0.0000263
Triiodo-L-Thyroxine
0.00000475-0.0000053
TWEEN 80 ®
0.002375-0.002625
Vascular Endothelial Growth Factor
0.0000046-0.00000525
Vitamin E
0.0019-0.0021.
15. The method of claim 1 , wherein the moiety of the nanoparticles of the composition that is capable of binding and releasing oxygen is a heme protein.
16. The method of claim 1 wherein said hydrophilic inner core of the nanoparticles contains a biologically active moiety.
17. The method of claim 16 wherein said biologically active moiety is selected from the group consisting of chemotherapeutic agents, proteins, peptides, polypeptides, enzymes and mixtures thereof.
18. The method of claim 1 wherein the composition comprises three phases: wherein the first and second phases are admixed with a third phase comprising a nanoparticle having an outer lipophilic coating and a hydrophilic inner core comprising a solution or suspension comprising a moiety capable of binding and releasing oxygen.
19. The method of claim 18 wherein the hydrophilic inner core of the third phase comprises a biologically active moiety.
20. The method of claim 1 conducted at a non-hypothermic temperature.
21. The method of claim 20 wherein the non-hypothermic temperature ranges from about 20 to about 37° C.
22. The method of claim 1 wherein the amount of each component of the composition is at the midpoint of the range expressed in the table.
23. The method of claim 1 wherein the mammalian heart is a human heart.