Crustacean Cell Culture Medium for Long-Term Viability and Proliferation
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Solution Overview
Problem
There is no continuous crustacean cell line available, and existing media for crustacean cell cultures do not support long-term viability and proliferation, with issues in osmolality, pH regulation, and nutrient composition leading to poor cell maintenance and growth.
Innovation Solution
A cell culture medium with a specific salt composition (sodium chloride, magnesium sulfate, magnesium chloride, calcium chloride, potassium chloride) to achieve an osmolality of 835 +/- 55 mmol/kg and pH of 7.2 to 7.4, supplemented with amino acids (glycine, L-alanine, L-aspartic acid, L-asparagine, L-glutamic acid, L-alanyl-L-glutamine, L-proline, L-serine, L-cysteine, L-tyrosine), vitamins (choline chloride, calcium pantothenate, folic acid, nicotinamide, pyridoxal hydrochloride, riboflavin, thiamine hydrochloride, inositol, biotin), glucose, lactalbumin, and ultrafiltered crustacean hemolymph to sustain and induce proliferation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If conventional media are used for crustacean cell cultures, then cell isolation and initial growth can be achieved, but long-term viability and proliferation are not supported
Solution Approach 1:
The patent optimizes multiple medium parameters including osmolality (adjusted to match crustacean hemolymph), pH buffering capacity, amino acid concentrations, and vitamin compositions. These parameter changes create an environment that supports both long-term cell viability and sustained proliferation, resolving the contradiction between duration and reliability.
Solution Approach 2:
The culture medium employs a composite formulation combining inorganic salts, organic compounds, amino acids, vitamins, and crustacean hemolymph components. This composite approach creates a synergistic effect that provides both the stability needed for long-term viability and the nutrients required for continuous proliferation.
2Reliability
If existing media compositions are used, then basic cell maintenance is possible, but correct osmolality and pH regulation fail
Solution Approach 1:
The patent specifically adjusts the osmolality of the culture medium to match that of crustacean hemolymph and implements a buffering system to maintain pH stability. These parameter changes ensure both reliable cell maintenance and stable chemical composition, eliminating the contradiction between maintenance quality and composition stability.
3Ease of manufacture
If primary cell cultures are attempted without optimized media, then initial cell isolation can occur, but subculturing and continuous proliferation fail
Solution Approach 1:
The patent prepares the culture medium in advance with all necessary nutrients, growth factors, and optimized conditions before cell isolation. This preliminary preparation ensures that when cells are isolated and subjected to subculturing, they immediately encounter an environment conducive to proliferation, thus maintaining both ease of isolation and high productivity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The medium maintains crustacean cells viable for extended periods and supports proliferation, enabling successful subculturing and proliferation, crucial for physiological studies and pathogen research.
Implementation Method 1
a specific salt composition, in part responsible for the correct osmolality and pH
Implementation Method 2
a specific salt composition, in part responsible for the correct osmolality and pH
Data Source
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AI summary
The present invention relates to a cell culture medium which sustains the growth of cells obtained from a crustacean in vitro. Indeed, the present invention discloses a cell culture medium comprising a specific salt composition, in part responsible for the correct osmolality and pH, a specific amino acid and vitamin composition, energy sources such as glucose, and crustacean hemolymph. The present invention further discloses that said cell culture medium is capable to keep cells obtained from shrimp and lobster viable for long periods of time even after passaging the cells into subcultures and induces said cells to proliferate. The cultured cells are for example useful to perform physiological studies on certain genes (by genetic engineering) and to do research and diagnosis on crustacean pathogens.