[go: up one dir, main page]

WO1989002919A1 - Procede de culture de perles - Google Patents

Procede de culture de perles Download PDF

Info

Publication number
WO1989002919A1
WO1989002919A1 PCT/US1988/003325 US8803325W WO8902919A1 WO 1989002919 A1 WO1989002919 A1 WO 1989002919A1 US 8803325 W US8803325 W US 8803325W WO 8902919 A1 WO8902919 A1 WO 8902919A1
Authority
WO
WIPO (PCT)
Prior art keywords
medium
source
nucleus
nacre
mollusca
Prior art date
Application number
PCT/US1988/003325
Other languages
English (en)
Inventor
Lee Dosuk
Original Assignee
Lee Dosuk
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Lee Dosuk filed Critical Lee Dosuk
Priority to KR1019890700947A priority Critical patent/KR890701006A/ko
Publication of WO1989002919A1 publication Critical patent/WO1989002919A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0601—Invertebrate cells or tissues, e.g. insect cells; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/32—Amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/38—Vitamins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/60—Buffer, e.g. pH regulation, osmotic pressure
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/70—Undefined extracts
    • C12N2500/74—Undefined extracts from fungi, e.g. yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/10—Mineral substrates
    • C12N2533/18—Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells

Definitions

  • the subject invention concerns coating of nucleating agents with nacre i_n vitro using mantle tisssue from mollusca.
  • This invention relates to a methods and compositions of matter utilized therein, for culturing pearls, and in particular to a method of producing pearls by using an in vitro cell culture system to maintain biological and physiochemical activities of the mantle tissues found in pearl forming mollusks and gastropods.
  • a pearl is a gem most commonly formed by bivalve mollusks, particularly by several marine species known as pearl oysters. Pearls are formed as a protection against the irritation caused by foreign objects, generally parasites, grains of sand, or bits of gravel, which lodge inside the shell. A fold of soft tissue envelopes the foreign particle and deposits layer after layer of nacre on it, similar to the mother-of-pearl lining the shell. Any shelled mollusk or gastropod is theoretically capable of producing a pearl, but most pearls exhibiting gem quality are produced by a specific genus of pinctada, haliotis, and certain freshwater bivalves.
  • Cultured pearls are generally produced by manually implanting a nucleus with small pieces of mantle tissue in an oyster for approximately two to three years.
  • the present invention circumvents the use of live oysters for culturing pearls. It is a process based on a cell culture system specifically designed and developed to provide sustenance and nourishment to pearl forming tissues kept in vitro. The process involves sustaining the viability of mantle tissue explants by providing necessary chemical and physical components in a culture medium.
  • the composition of the medium to maintain and grow tissues includes a combination of nutrients associated with the naturally available medium in which the tissue grows, which medium may include inorganic salts, a source of amino acids, a sugar source, naturally occurring proteins, etc.
  • mantle tissue when mantle tissue is extracted from a mollusk and maintained under certain _in vitro conditions, the tissue has physiological characteristics and functions duplicating those in normal biological conditions.
  • the present invention uses complex growth media with the appropriate chemical and physical components necessary to simulate the ideal conditions in vitro for mantle tissues.
  • the chemical and ultrastructural properties of the pearls produced by this invention are substantially identical to the naturally occurring or cultured pearls currently on the market, as has been demonstrated by a series of electron micrographs comparing the ultrastructure and chemical properties of conventionally cultured pearls with those of the pearls produced by the present invention. From transmission electron micrographs illustrating the surface structure of a conventionally produced cultured pearl presently found on the market and a pearl produced from the present invention, it could be seen that crystal size and shape of the nacre as well as general surface topography are substantially identical between the two pearls. Chemical identifications using X-ray diffraction of a conventionally produced cultured pearl and a pearl produced by the present invention showed that calcium carbonate (aragonite) is the predominate phase found in both pearls.
  • the culture media used in the process of this invention are complex growth media.
  • the media employed may take many forms, normally providing an environment which to varying degrees approaches the natural fluids, cellular or extracellular.
  • the medium will include inorganic salts, a source of amino acids, a source of metabolic energy, normally a metabolizable or assimilable saccharide source, and desirably a source of hormones.
  • the various components of the nutrient medium which may be involved may be separated generally into the following components: (i) inorganic salts, (ii) amino acids, (iii) vitamins, (iv) naturally occurring protein containing compositions, (v) antibiotics and antimycotics, (vi) assimilable energy source and (vii) C0 2 , CO3 and/or 0 2 .
  • the nutrient medium will have the inorganic salts necessary for growth of the tissue, approximating the natural medium.
  • various other factors may be included, where some factors may be provided by the presence of other factors.
  • the naturally occurring material may include the factors which are otherwise added to a synthetic medium.
  • the naturally occurring material may serve as an alternative to another component. Therefore, there may be wide variation in the media which are employed, and a number of different media may be employed, each with different degrees of success in the rapidity with which the pearls are obtained.
  • the tissue when using mollusk tissue as a component of the medium, the tissue may serve as a source of amino acids and hormones. Protein hydrolysates may serve as a source of amino acids, so that individual amino acids may not be included.
  • Conditioned medium from the growth of mantle tissue may be employed, where the conditioned medium may be replenished to previous levels of certain components such as an energy source and undesirable factors reduced to a desirable level.
  • the basic medium will have a salt medium approximating the natural medium of the tissue at an appropriate pH, a source of assimilable energy, a source of essential amino acids, and additional factors which promote growth.
  • factors may include mollusca tissue, particularly the gonadal tissue components, which may also serve as the source of amino acids, fetal serum, a steriod source such as an androgen extract, yeast extract or the like.
  • the inorganic salts in the growth medium will usually contain all of the ions that are found in the mollusk mantle tissues themselves during their biological activities. In order to determine the chemical nature of the fluid surrounding the mantle tissue, the ionic concentrations of such fluid were investigated and determined. In Table I, the average concentrations of inorganic elements found are shown.
  • compositions of inorganic salts were formulated for use in the complex growth medium as found in Table II below.
  • salt media proposed above are convenient, but other salt media may be formulated which would provide the desired ionic composition and normalities of the essential ions.
  • L-valine Of particular interest as a source of individual amino acids are the amino acids serine, alanine, arginine, aspartic acid, glycine, histidine, leucine, methionine, and phenylalanine.
  • amino acids it is understood that the naturally occurring L-stereoisomer is intended.
  • protein hydrolysates or extracts may be employed, by themselves or in conjunction with individual amino acids, where the protein hydrolysates or extracts may provide for a more economical source of amino acids.
  • Vitamins are optional. If included, the vitamins present in the complex growth medium are similar to those used for other cell and tissue cultures with some variations in concentration. Specific vitamins and concentrations are shown in Table V below. None some or all may actually be used in the complex growth medium.
  • Vitamin A (acetate)
  • the hormones found associated at or near the activities of the mantle tissues were determined and may be classified as corticosteroids, androgens or modified versions of estrogen. These hormonal components or metabolic intermediates were added to the medium to facilitate biosynthesis of these hormones.
  • Androgen extract may be obtained from commercial sources and may be used in amounts varying from 0.005 to 0.2 mg/L. It has been observed that a growth medium containing serum extracts from the reproductive organs of the same mollusk was observed to yield the most success in proliferating the growth of the mantle tissues. The serum extracts were found to provide the levels of hormones necessary to sustain mantle tissue growth. Without the presence of this reproductive organ serum, however, the hormone additives, although not essential in maintaining tissue culture, were found to be desirable in enhancing tissue growth.
  • tissue from the mantle explant source or other related species particularly comprising gonadal tissue or lyophilized gonadal tissue.
  • the tissue may be minced or ground either before or after lyophilization to provide the tissue in easily dispersible form, e.g. a powder.
  • the tissue when employed will be composed of cells, fragments of the cells and other materials present in the tissue when removed from the source.
  • the "lyophilized tissue may be dispersed in the nutrient medium under mild conditions with agitation, and may be incorporated in biocompatible gels, such as agar or collagen gels.
  • the amount of the lyophilized tissue may be varied widely and may be optimized in accordance with the other components of the nutrient medium. Beneficial results may be obtained by using a ratio of from about 0.1 to 2 of the volume of the tissue prior to lyophilization to the volume of medium.
  • Antibiotics and anti ycotics such as penicillin and streptomycin may also be used during tissue extraction procedures and may also be used to control contamination levels during mantle tissue culture.
  • the antibiotics and antimycotics used in the complex growth medium are listed below in Table VI.
  • Saccharide sources include glucose, galactose, glycogen, sucrose, etc.
  • lactalbumin hydrolysate provided significant benefits in the absence of other protein sources.
  • the system is buffered to provide a physiologically acceptable pH.
  • Sodium bicarbonate (inorganic salt) and phenol red were added to control and monitor the pH of the solution.
  • the preferred pH range is 7.2 to 7.9.
  • a pH as low as 6.5 and as high as 9.3 have been observed to sustain viability of the mantle tissues.
  • Carbon dioxide enriched air or a carbonated source was added to the growth medium to keep the pH of the medium within the overall range of 6.6 to 8.9.
  • Vitamin A (acetate)
  • Formulations were prepared for growth media using distilled water containing the salt composition of Medium A or sea water and gonad organ extracts, where the gonad organ extracts were obtained by isolating the gonads from the body mass of the mollusc, lyophilizing the gonadal tissue and grinding the tissue to form a powder. The powder was then uniformly dispersed in the water by stirring. The amount of gonadal tissue was based on the original volume of the tissue and was in a volume ratio to water of 0.25 to 1. Other formulations combined the above formulation or used androgen extract (Sigma) in place of the gonadal tissue. The androgen extract ranged in the amount of 0.01 to O.lmg/L.
  • a portion or all of a mantle tissue is explanted from a mollusk or gastropod which belong to genus thereof which produce mother-of-pearl material in their shells, such as the genera Pinctada, Isognomon, Pteria, Pinna, Haliotis, Atrima, etc., e.g., Pinctada martensii, Pinctada margritifera, Pinctada maxima, Pinctada fucata, Pteria macroptera, Atrima japonica, Ostrea gigas, Unio margritifera, Cristaria plicata, Tridacna gigas, Haliotis gigantea, ⁇ ypriopsis schlegeli, Haliotis fulgens, Haliotis corrugata, Haliotis cracherodii, Haliotis discus hannai, Ligumia n
  • the pieces of the mantle tissue fragments are optionally washed in seawater containing antibiotics and/or antimycotics, e.g., 100 Units penicillin G (sodium salt)/ml and 100 ug/ml streptomycin.
  • the seawater may be prepared in the laboratory and would consist generally of water with inorganic salts in concentrations thereof as indicated in Table II above.
  • the tissue part(s) or intact mantle tissue(s) are transferred into a carbon dioxide or oxygen enriched complex growth medium comprised of inorganic salts, amino acids, vitamins and growth factors, hormones, antibiotics and antimycotics, animal serum extracts and glucose.
  • Biocompatible material such as glass, porcelain, old shells containing calcite and/or the aragonite phase(s) of calcium carbonate, and/or calcium phosphates in the form of spheres or hemispheres are introduced into the medium for nacre deposition.
  • the material which serves as a nucleating substrate may be of any shape, normally being round or ellipsoid for pearl formation.
  • the smallest dimension of the material which serves as the substrate will be usually at least about 1mm, usually at least about 2mm and may be as large as 20mm or larger.
  • the substrate will usually have a diameter from about 2 to 15, usually 7 to 10mm.
  • small, i.e., less than 1 m ⁇ r pieces of mantle tissue were sectioned and removed from a pearl forming adult mollusk. Each piece was washed in saltwater containing the inorganic salt components described above and placed in pre-sterilized culture dishes. Sterilized complex growth medium, filtered through a 0.22 ⁇ m millipore filter (Millipore Corp., Bedford, MA 01730) was immediately added to each dish containing the tissue fragments. Small nuclei, i.e., glass beads or spheres made from clam shells, were then placed in each dish and positioned so that maximal physical contact was made with the tissue.
  • the medium is changed every two days or when the medium pH rises above 7.8, whichever occurs sooner and depending on tissue mass to medium content ratio.
  • Precise measurement of pH may be done with a pH measuring apparatus. After the pearl reached its desired quality and/or size, it was removed from the dish and the attached tissue removed. The removed tissue was dissected and fragments thereof used again in the production of additional pearls.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Environmental Sciences (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Farming Of Fish And Shellfish (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'enrobage de nacre d'un substrat de nucléation biocompatible est réalisé par mise en contact d'un tel substrat avec un explant tissulaire de couverture provenant d'espèces mollusques dans des conditions favorisant la croissance cellulaire. On utilise des milieux contenant du sel, avec des variations quant aux autres additifs, pouvant comprendre une source de matière stéroïde, une source d'acides aminés, une source d'énergie métabolisable, ou d'autres composants. Les substrats enrobés de nacre sont formés après incubation pendant une durée suffisante. Par un choix approprié des noyaux on peut produire des perles.
PCT/US1988/003325 1987-09-28 1988-09-27 Procede de culture de perles WO1989002919A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1019890700947A KR890701006A (ko) 1987-09-28 1988-09-27 진주양식법

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US10212787A 1987-09-28 1987-09-28
US102,127 1987-09-28

Publications (1)

Publication Number Publication Date
WO1989002919A1 true WO1989002919A1 (fr) 1989-04-06

Family

ID=22288251

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1988/003325 WO1989002919A1 (fr) 1987-09-28 1988-09-27 Procede de culture de perles

Country Status (6)

Country Link
EP (1) EP0335955A4 (fr)
JP (1) JPH02501441A (fr)
KR (1) KR890701006A (fr)
AU (1) AU611508B2 (fr)
CA (1) CA1321964C (fr)
WO (1) WO1989002919A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993008265A2 (fr) * 1991-10-23 1993-04-29 Georges Camprasse Productions d'os et de nacre, a partir de cellules formatrices d'os mises en presence de nacre
WO1993011224A1 (fr) * 1991-12-06 1993-06-10 Bertin & Cie Societe Anonyme Milieux de culture pour des cellules d'invertebres marins
US7062940B2 (en) 2002-12-13 2006-06-20 Chi Huynh Carved pearl
EP2084286A1 (fr) * 2006-11-22 2009-08-05 Indian Council Of Agricultural Research Production de perles in-vitro à l'aide d'organismes marins
CN112919943A (zh) * 2021-03-26 2021-06-08 生态环境部华南环境科学研究所 一种高效去除畜禽粪便中类固醇类雌激素的堆肥调剂及好氧堆肥方法

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110637763B (zh) * 2019-09-26 2021-05-04 中国科学院南海海洋研究所 一种砗磲贝壳形态与外套膜颜色性状互换品系的制备方法
CN110684709B (zh) * 2019-10-21 2021-01-22 福建罗屿岛食品有限公司 一种皱纹盘鲍细胞培养基及细胞系的构建方法

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
C R Hebd Seances Acad Sci Ser D Sci Nat 278 (19), 1974, HOUTTEVILLE et al, "Experimental Analysis in Organ Culture of the Action of Cerebro Pleural and Visceral Ganglia on the Mantle of the Male Mussel Mytilus-Edulis Mollusca Pelecypoda", pages 2469-2472, title only. *
Grant, "Hackh's Chemical Dictionary" 1972 McGRAW-HILL Book Company, New York pages 493. *
Mem Fac Educ Shiga Univ Nat Sci 1985, HIGASHI et al, "Organ Culture of Mantle Tissue of the Freshwater Mussel Hyriopsis-Schlegeli", pages 39-44, title only. *
Rechcigl, "CRC Handbook Series in Nutrition and Food, Section G Volume IV, Culture Media for Cells, Organs and Embryos" 1977, CRC Press, Cleveland, Ohio, pages 131, 132, and 169. *
See also references of EP0335955A4 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993008265A2 (fr) * 1991-10-23 1993-04-29 Georges Camprasse Productions d'os et de nacre, a partir de cellules formatrices d'os mises en presence de nacre
FR2682965A1 (fr) * 1991-10-23 1993-04-30 Camprasse Georges Production d'os et de perle a partir de culture de cellules formatrices d'os.
WO1993008265A3 (fr) * 1991-10-23 1993-09-16 Georges Camprasse Productions d'os et de nacre, a partir de cellules formatrices d'os mises en presence de nacre
WO1993011224A1 (fr) * 1991-12-06 1993-06-10 Bertin & Cie Societe Anonyme Milieux de culture pour des cellules d'invertebres marins
FR2684686A1 (fr) * 1991-12-06 1993-06-11 Bertin & Cie Milieux de culture pour des cellules d'invertebres marins.
US7062940B2 (en) 2002-12-13 2006-06-20 Chi Huynh Carved pearl
EP2084286A1 (fr) * 2006-11-22 2009-08-05 Indian Council Of Agricultural Research Production de perles in-vitro à l'aide d'organismes marins
EP2084286A4 (fr) * 2006-11-22 2011-11-23 Indian Agricultural Council Production de perles in-vitro à l'aide d'organismes marins
CN112919943A (zh) * 2021-03-26 2021-06-08 生态环境部华南环境科学研究所 一种高效去除畜禽粪便中类固醇类雌激素的堆肥调剂及好氧堆肥方法

Also Published As

Publication number Publication date
AU2608188A (en) 1989-04-18
EP0335955A1 (fr) 1989-10-11
KR890701006A (ko) 1989-12-19
JPH02501441A (ja) 1990-05-24
CA1321964C (fr) 1993-09-07
EP0335955A4 (fr) 1990-05-14
AU611508B2 (en) 1991-06-13

Similar Documents

Publication Publication Date Title
Ebner et al. Cultivation and properties of bovine mammary cell cultures
Rivkin et al. Bacterivory: a novel feeding mode for asteroid larvae
Gallardo et al. Effect of some vertebrate and invertebrate hormones on the population growth, mictic female production, and body size of the marine rotifer Brachionus plicatilis Müller
Ambesi-Impiombato et al. Thyroid cells in culture
AU724447B2 (en) Medium composition for external fertilization
CN101491228A (zh) 一种培育海水无核珍珠的方法
Desrosiers et al. Early developmental events following fertilization in the giant scallop () Placopecten magellanicus
Cheng et al. Aquaculture of the tropical sea cucumber, Stichopus monotuberculatus: Induced spawning, detailed records of gonadal and embryonic development, and improvements in larval breeding by digestive enzyme supply in diet
CA2537462A1 (fr) Milieu de culture cellulaire comprenant du fructose en tant que source d'energie principale
Fraser et al. Studies on primary cell cultures derived from ovarian tissue of Penaeus monodon
Gong et al. Characterization of calcium deposition and shell matrix protein secretion in primary mantle tissue culture from the marine pearl oyster Pinctada fucata
CA1321964C (fr) Methode de culture perliere
Sud et al. Role of water-soluble matrix fraction, extracted from the nacre of Pinctada maxima, in the regulation of cell activity in abalone mantle cell culture (Haliotis tuberculata)
Wyss Ecdysterone, insulin and fly extract needed for the proliferation of normal Drosophila cells in defined medium
Xing et al. The potential value of different species of benthic diatoms as food for newly metamorphosed sea urchin Strongylocentrotus intermedius
US4449480A (en) Culture of freshwater mussel glochidia in an artificial habitat utilizing complex liquid growth media
CN108265021B (zh) 一种水蛭细胞体外培养基及其培养方法
Mercurio et al. Primary cell cultures from sea urchin ovaries: a new experimental tool
Dautov et al. Towards a scientific-based farming of sea urchins: First steps in the cultivation of Diadema setosum, Diadema savignyi and Mesocentrotus nudus
Le Marrec-Croq et al. Primary cultures of heart cells from the scallp Pecten maximus (mollusca-bivalvia)
JPS60114192A (ja) 蟻毒腺細胞の培養方法及び培地
Kirk et al. Morphologically stable epithelial vesicles cultured from normal human endometrium in defined media
Ferkovich et al. Rearing of ectoparasitoid Diapetimorpha introita on an artificial diet: supplementation with insect cell line-derived factors
Stanley Cultivation of arthropod cells
CN109022346B (zh) 一种海洋无脊椎动物细胞培养基

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU JP KR

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LU NL SE

WWE Wipo information: entry into national phase

Ref document number: 1988909428

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1988909428

Country of ref document: EP

WWR Wipo information: refused in national office

Ref document number: 1988909428

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1988909428

Country of ref document: EP