Chemistry: molecular biology and microbiology – Maintaining blood or sperm in a physiologically active state...
Patent
1997-07-15
1999-02-02
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Maintaining blood or sperm in a physiologically active state...
435 41, 435 711, 435 696, 435267, 435269, 424538, A01N 102, C12P 2104, A61K 3564
Patent
active
058663177
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a method for collecting hemolymph of insects while making the most use of the self-contraction phenomenon caused in response to the freezing and thawing operations of larvae of insects, in particular, lepidopterous insects.
BACKGROUND ART
The hemolymph of insects belonging to, for instance, the lepidopteran has conventionally been collected manually by partially puncturing the epidermis of the insect bodies and then squeezing the hemolymph out of the insect bodies. For this reason, the operations for collecting hemolymph, at a time, from a vast number of insect bodies requires much labor and a great deal of time and are thus impracticable. Moreover, the hemolymph may be scattered during squeezing the same out of the insect bodies.
Moreover, in respect of the collection of the hemolymph from larvae of silkworm moth, there has been proposed a method for incising silkworms using a blade which can emit ultrasonics (Japanese Un-Examined Patent Publication No. Hei 9-19238). However, such an incision method requires the use of special equipments and apparatuses and has not widely been used. In addition, such a method has not been applied, at all, to the collection of hemolymph from other insects such as lepidopterous insects.
DISCLOSURE OF THE INVENTION
The foregoing conventional techniques are not satisfied since they are not a method for efficiently collecting hemolymph from insects such as those belonging to the lepidopteran and accordingly, the following problems should be solved: vast number of insect bodies within a short period of time; collection thereof; equipment and apparatus and can widely be used.
Further, the foregoing methods would suffer from a problem in that the hemolymph may be contaminated with unnecessary tissues or the like originated from the epidermis of the insects incised for the extraction of the hemolymph. Therefore, it is desirable that the opening formed through incision to extract the hemolymph has the smallest necessary size. Moreover, the insect hemolymph externally extracted from the insect bodies undergoes melanization and therefore, the resulting hemolymph must be subjected to a melanization-inhibitory treatment immediately after the extraction thereof. Accordingly, the hemolymph-collection method should further satisfy the following requirements: the like into the extracted hemolymph; and hemolymph.
Accordingly, an object of the present invention is to eliminate the foregoing problems and to provide a method for collecting insect hemolymph which permits the collection of hemolymph, at a time, from a vast number of insect bodies, which does not cause any scattering of the hemolymph during collection thereof, which may widely be used, which permits the collection of the hemolymph substantially free of unnecessary tissues or the like and which can inhibit the melanization of the collected hemolymph.
Baculovirus-expressing cell lines obtained using Bombyx mori larvae as host cells among the lepidopterous insects can express extremely large amount of extraneous genes and thus may serve as a means for producing a large amount of a recombinant useful protein. For this reason, a method for efficiently collecting insect hemolymph developed for the extraction of the protein expressed in a large amount within silkworm bodies closely relates to the development of novel technique for the production of rare and useful proteins. In addition, the insect hemolymph is often used as a culture medium for the cultivation of insect cells. For this reason, such a method for efficient collection of insect hemolymph would widely be used not only in the industrial fields, but also in researches which make use of the insect hemolymph. Under such circumstances, the inventors of this invention have conducted intensive studies to develop a method for efficient collection of insect hemolymph and have thus completed the present invention.
The method for collecting insect hemolymph according to the present invention comprises the steps of freezing anesth
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Asako Terasaki et al. "Effect of silkworm hemolymph treated with sodium thiosulphate on the growth of insect cell cultures", J. Sericultural Science of Japan 1990, 59(5), 350-354 & BIOSIS Abstract No. 91037564.
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Arakawa Toru
Miyazawa Mitsuhiro
Japan as Represented by Director General of National Institute o
Lankford , Jr. Leon B.
Tate Christopher R.
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