In vitro cultivation of horseshoe crab amebocytes
AssigneeBaxter Travenol Laboratories, Inc.
InventorFrederick C. Pearson
Life of this patent
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Abstract
A culture medium for cultivating horseshoe crab amebocytes, called Limulus amebocytes, composed of inorganic salts, vitamins, amino acids, and carbon sources including glucose is given. The process for cultivating the Limulus amebocytes depends on maintaining a pH between 6.5 and 9.0 while agitating the medium. Production of Limulus lysate from these cells is an eventual goal.
Claims4
A culture medium for cultivating Limulus amebocytes, comprising glucose, L-cystine, salts, vitamins, amino acids and horseshoe crab serum.
A culture medium for cultivating Limulus amebocytes in vitro comprising horseshoe crab serum and:
- NaCl:
- 1000 mg/L
- CaCl.sub.2 :
- 30.0
- KCl:
- 30.0
- MgCl.sub.2.6H.sub.2 O:
- 2280.
- MgSO.sub.4.7H.sub.2 O:
- 2780.
- NaHCO.sub.3 :
- 350
- NaH.sub.2 PO.sub.4.H.sub.2 O:
- 1013.0
- alpha Ketoglutaric acid:
- 370.
- Glucose:
- 1000.
- Malic acid:
- 670
- L-alanine:
- 225
- L-Arginine HCl:
- 700.0
- L-Asparagine:
- 350.00
- L-Aspartic acid:
- 350.0
- L-Cystine:
- 100.
- L-Glutamic acid:
- 600.00
- L-Glutamine:
- 600.00
- Glycine:
- 650.00
- L-Histidine:
- 2500.00
- L-Isoleucine:
- 50.00
- L-Leucine:
- 75.00
- L-Lysine HCl:
- 625.00
- L-Methionine:
- 50.00
- L-Phenylalinine:
- 150.00
- L-Proline:
- 350.00
- DL-Serine:
- 1100.00
- L-Threonine:
- 175.00
- L-Tryptophane:
- 100.00
- L-Tyrosine:
- 50.00
- L-Valine:
- 100.0
- VITAMINS
- Biotin:
- 0.01
- D-Ca pantothenate:
- 0.02
- Choline chloride:
- 0.20
- Folic acid:
- 0.02
- i-Inositol:
- 0.02
- Niacin:
- 0.02
- Para-aminobenzoic acid:
- 0.02
- Pyridoxine HCl:
- 0.02
- Riboflavin:
- 0.02
- Thiamine HCl:
- 0.02.
A process for propagating Limulus amebocytes in vitro, comprising inoculating Limulus amebocytes into a nutrient medium comprising a carbon source, salts, vitamins, amino acids and horseshoe crab serum, constantly agitating the suspension of amebocytes in the medium at a temperature of from 12.degree. C. to 30.degree. C., and isolating the amebocytes from the suspension.
Dependencies are inferred from the claim text.