Liquid crystals with reduced toxicity and applications thereof

• US 20050079487A1
• Filed: 07/16/2004
• Published: 04/14/2005
• Est. Priority Date: 07/17/2003
• Status: Active Grant

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Abbott, Nicholas L., Jang, Chang-Hyun, Murphy, Christopher John, Campbell, Sean Francis, Cheng, Li-Lin, Luk, Yan-Yeung
• Granted Patent: US 7,303,694 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07D 311/24 with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
• CPC Class Codes: C09K 19/3003 Compounds containing at least two rings in which the different rings are directly linked (covalent bond)
• CPC Class Codes: C09K 19/42 Mixtures of liquid crystal compounds covered by two or more of the preceding groups C09K19/06 - C09K19/40
• CPC Class Codes: C09K 2019/0407 containing a carbocyclic ring, e.g. dicyano-benzene, chlorofluoro-benzene or cyclohexanone
• CPC Class Codes: C09K 2019/301 Cy-Cy-Ph

Abstract

Liquid crystal compositions that exhibit little or no toxicity with respect to cells include liquid crystals with chemical functional groups such as fluorine atoms, fluorophenyl groups, or difluorophenyl groups. Liquid crystals with little or no toxicity to cell lines may be added to cell culture media or added to components used in cell culture media. Cells may be grown in cell culture media that includes liquid crystals that exhibit little or no toxicity to cells.

Claims

1. A liquid crystal composition, comprising:

at least two different liquid crystal compounds, wherein the first liquid crystal and the second liquid crystal both comprise at least one fluorine group, and at least one of the first liquid crystal and the second liquid crystal comprises a fluorinated phenyl group.

2. The liquid crystal composition of claim 1, wherein the fluorinated phenyl group is a difluorinated phenyl group.

3. The liquid crystal composition of claim 1, wherein at least one of the first liquid crystal or the second liquid crystal is a compound of the following formula wherein Z is a methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, or hexadecyl group

4. The liquid crystal composition of claim 1, wherein both the first liquid crystal and the second liquid crystal are compounds of the following formula, wherein Z is a propyl group in the first liquid crystal and Z is a pentyl group in the second liquid crystal

5. The liquid crystal composition of claim 1, wherein the first liquid crystal is 4′

-(3,4-difluorophenyl)-4-propylbicyclohexyl and the second liquid crystal is 4′

-(3,4-difluorophenyl)-4-pentylbicyclohexyl and the molar ratio of the first liquid crystal to the second liquid crystal ranges from 10;

90 to 90;

10.

6. The liquid crystal composition of claim 5, wherein the molar ratio of the first liquid crystal to the second liquid crystal ranges from 45:

55 to 55;

45.

7. The liquid crystal composition of claim 1, further comprising at least one cell culture medium component selected from a vitamin, an amino acid, a growth factor, or combinations of these.

8. The liquid crystal composition of claim 1, wherein the liquid crystal composition comprises at least one vitamin and at least one amino acid.

9. The liquid crystal composition of claim 1, further comprising a cell and at least one cell culture medium component selected from a vitamin, an amino acid, a growth factor, or combinations thereof.

10. The liquid crystal composition of claim 9, wherein the cell is a stem cell, a 3T3 fibroblast, or a SV-40 transformed human corneal epithelial cell.

11. The liquid crystal composition of claim 9, wherein the cell is eukaryotic or prokaryotic cell.

12. The liquid crystal composition of claim 10, wherein the cell is selected from a vertebrate cell, or an invertebrate cell.

13. The liquid crystal composition of claim 12, wherein the vertebrate cell is selected from a fish cell, an amphibian cell, a reptile cell, a bird cell, or a mammal cell, and the invertebrate cell is selected from an annelid cell, a mollusk cell, or an insect cell.

14. The liquid crystal composition of claim 9, wherein the cell is a human cell.

15. The liquid crystal composition of claim 9, wherein the cell is a mouse, rat, hamster, guinea pig, monkey, ape, cat, dog, horse, pig, cow, plant, bacteria, or mycoplasmal cell.

16. A liquid crystal cell culture media, comprising at least one liquid crystal compound and at least one cell culture component selected from a vitamin, an amino acid, a sugar, or combinations thereof, wherein the at least one liquid crystal includes at least one fluorine group, is a cholesteric liquid crystal, or is a liquid crystal that exhibits reduced toxicity towards cells.

17. The liquid crystal cell culture media of claim 16, wherein the cell culture media comprises at least three amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

18. The liquid crystal cell culture media of claim 17, wherein the cell culture media comprises at least five amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

19. The liquid crystal cell culture media of claim 18, wherein the cell culture media comprises at least eight amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

20. The liquid crystal cell culture media of claim 16, wherein the cell culture media further comprises at least two salts selected from CaCl₂, NaCl, NaHCO₃, Na₂HPO₄, NaH₂PO₄, KCl, K₂HPO₄, KH₂PO₄, MgSO₄, MgCl₂, Fe(NO₃)₃, FeSO₄, CuSO₄, ZnSO₄, ZnCl₂, COCl₂, CuCl₂, MnCl₂, (NH₄)₂MoO₄, or hydrates or combinations thereof.

21. The liquid crystal cell culture media of claim 16, wherein the cell culture media comprises at least two vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

22. The liquid crystal cell culture media of claim 21, wherein the cell culture media comprises at least five vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

23. The liquid crystal cell culture media of claim 22, wherein the cell culture media comprises at least eight vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

24. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal is a cholesteric liquid crystal.

25. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal comprises a fluorinated cyclohexyl group.

26. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal comprises a fluorinated phenyl group.

27. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal comprises a difluorinated phenyl group.

28. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal comprises a difluorinated compound having the following formula where Z is an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 carbon atoms

29. The liquid crystal cell culture media of claim 16, wherein the at least one liquid crystal is a lyotropic or a thermotropic liquid crystal.

30. The liquid crystal cell culture media of claim 16, wherein the liquid crystal cell culture media comprises 2, 3, 4, 5, or more different liquid crystals.

31. The liquid crystal cell culture media of claim 16, further comprising water.

32. The liquid crystal cell culture media of claim 16, further comprising glucose.

33. A kit for growing cells, comprising:

(a) at least one liquid crystal selected from a cholesteric liquid crystal, a liquid crystal that comprises at least one fluorine atom, or a liquid crystal that exhibit reduced toxicity towards cells; and

(b) a cell culture medium comprising at least one cell culture component selected from a vitamin, an amino acid, a sugar, or combinations thereof.

34. The kit of claim 33, wherein the cell culture medium comprises at least three amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

35. The kit of claim 34, wherein the cell culture medium comprises at least five amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

36. The kit of claim 35, wherein the cell culture medium comprises at least eight amino acids selected from L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, glycine, L-histidine, hydroxy-L-proline, L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine, L-phenylalanine, L-proline, L-serine, L-taurine, L-threonine, L-tryptophan, L-tyrosine, L-valine, or salts or combinations thereof.

37. The kit of claim 33, wherein the cell culture medium comprises at least two salts selected from CaCl₂, NaCl, NaHCO₃, Na₂HPO₄, NaH₂PO₄, KCl, K₂HPO₄, KH₂PO₄, MgSO₄, MgCl₂, Fe(NO₃)₃, FeSO₄, CuSO₄, ZnSO₄, ZnCl₂, CoCl₂, CuCl₂, MnCl₂, (NH₄)₂MoO₄, or hydrates or combinations thereof.

38. The kit of claim 33, wherein the cell culture medium comprises at least two vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

39. The kit of claim 38, wherein the cell culture medium comprises at least five vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

40. The kit of claim 39, wherein the cell culture medium comprises at least eight vitamins selected from ascorbic acid, D-biotin, choline, choline chloride, choline bitartrate, folic acid, myo-inositol, inositol, niacin, niacinamide, nicotinamide, p-aminobenzoic acid, D-pantothenic acid, pyridoxine, pyridoxal, riboflavin, DL-thioctic acid, thiamine, vitamin B12, vitamin A alcohol, vitamin D-2, vitamin E, menadione, nicotinic acid, alpha-tocopherol, or salts or combinations thereof.

41. The kit of claim 33, wherein the at least one liquid crystal is a cholesteric liquid crystal.

42. The kit of claim 33, wherein the at least one liquid crystal comprises a fluorinated cyclohexyl group.

43. The kit of claim 33, wherein the at least one liquid crystal comprises a fluorinated phenyl group.

44. The kit of claim 33, wherein the at least one liquid crystal comprises a difluorinated phenyl group.

45. The kit of claim 33, wherein the at least one liquid crystal comprises a difluorinated compound having the following formula, wherein Z is an alkyl group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 carbon atoms

46. The kit of claim 33, wherein the at least one liquid crystal is a lyotropic liquid crystal.

47. The kit of claim 33, wherein the at least one liquid crystal is a thermotropic liquid crystal.

48. The kit of claim 33, wherein the kit comprises 2, 3, 4, 5, or more different liquid crystals.

49. The kit of claim 33, wherein the cell culture medium further comprises glucose.

50. The kit of claim 33, wherein the kit further comprises at least one support for cells.

51. The kit of claim 50, wherein the at least one support is a grid.

52. The kit of claim 50, wherein the at least one support is permeable.

53. The kit of claim 50, wherein the at least one support comprises a surface with topographic features that align liquid crystals on which cells may be placed.

54. The kit of claim 50, wherein the at least one support comprises a nanostructured surface to microstructured surface possessing features on which cells may be placed.

55. The kit of claim 53, wherein the features comprise peaks, ridges, columnar projections, pillars, or combinations thereof.

56. The kit of claim 33, wherein the kit further comprises instructions for growing cells in the cell culture medium in the presence of the at least one liquid crystal.

57. A composition, comprising:

a virus or a cell, a compound of formula I or a salt thereof, and a cell culture component selected from a vitamin, an amino acid, a sugar, or combinations thereof, wherein the compound of formula I has the following formula

58. The composition of claim 57, wherein the composition comprises the virus and the cell.

59. The composition of claim 57, wherein the virus is vesicular stomatitis virus.

60. The composition of claim 57, wherein the compound of formula I or the salt thereof is disodium chromoglycate.

61. The composition of claim 57, further comprising a poly-L-lysine treated coverslip.

62. A method for investigating viruses, comprising contacting a virus with a compound of formula I or a salt thereof, wherein the compound of formula I has the following formula

63. The method of claim 62, further comprising contacting the virus with the compound of formula I or the salt thereof in the presence of a cell.

64. The method of claim 63, further comprising contacting the virus with the compound of formula I or the salt thereof in the presence of the cell and a cell culture component selected from a vitamin, an amino acid, a sugar, or combinations thereof.

65. A method for detecting interactions between a biomolecule and a receptor, comprising:

(a) contacting an aqueous solution comprising water, a liquid crystal, and a biomolecule with a surface that uniformly orients the liquid crystal when the biomolecule does not bind to a receptor, wherein the receptor is either bound to the surface or is included in the aqueous solution, further wherein the surface resists adsorption of the biomolecule if the biomolecule does not bind to the receptor, but the biomolecule will be bound to the surface if the receptor is bound to the surface and the receptor binds the biomolecule, further wherein if the biomolecule binds to the receptor in the absence of the liquid crystal, then the biomolecule will still bind to the receptor in the presence of the liquid crystal.

66. The method of claim 65, wherein the biomolecule is selected from a peptide, a polypeptide, DNA, RNA, a DNA fragment, a RNA fragment, a cell, a virus, or a bacterium.

67. The method of claim 65, wherein the liquid crystal is a compound of formula I or a salt thereof, wherein the compound of formula I has the following formula

68. The method of claim 67, wherein the liquid crystal is disodium chromoglycate.

69. The method of claim 65, wherein the surface comprises a serum albumin, and the serum albumin resists adsorption of biomolecules that do not bind to the receptor.

70. The method of claim 65, wherein the receptor is included in the aqueous solution.

71. The method of claim 65, wherein the receptor is bound to the surface.

72. The method of claim 65, wherein the surface comprises a self-assembled monolayer on a metallized surface.

73. The method of claim 72, wherein the self-assembled monolayer is formed from a thiol having the formula HS—

(CH₂)_p—

(OCH₂CH₂)_q—

OH, wherein p is an integer with a value of from 5 to 20 and q is an integer with a value of from 1 to 6.

74. The method of claim 73, wherein p is 11 and q is 3 or 4.

75. The method of claim 65, wherein the surface comprises a glass surface that has been reacted with a compound of formula (R^aO)₃Si—

(CH₂)_s—

N═

C═

O and has then been reacted with the receptor, wherein Ra is a straight or branched chain alkyl group having 1 to 6 carbon atoms and s is an integer having a value of from 2 to 6.

76. The method of claim 75, wherein R^a is a methyl, ethyl, or propyl group and s is 3.

77. The method of claim 75, wherein the receptor is a protein.