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1991 Fiscal Year Final Research Report Summary

The Development of the Methods for Mass Culture, Maintenance and Transplantation of Various Cells Using Extracellular Matrix

Research Project

Project/Area Number 02554026
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 動物形態・分類学
Research InstitutionDokkyo University School of Medicine

Principal Investigator

YOMAOKA Sadao  DOKKYO UNIV., ECH. MED. Dept. Physiol., Prof., 医学部, 教授 (50049813)

Co-Investigator(Kenkyū-buntansha) WATANABE Kazuto  DOKKYO UNIV., SCH. MED., Dept. Physiol. Assist. Professor, 医学部, 助手 (80146167)
ENAMI Jumpei  ZENYAKU KOGYO Co, LTD., Dept. Biotech. Res. Lab., Manager, 研究所バイオテク研究室, 室長 (30112634)
KOHMOTO Kaoru  UNIV. TOKYO, FAC. AGR. Dept. Animal Breeding, Prof., 農学部, 教授 (30011894)
Project Period (FY) 1990 – 1991
KeywordsCollagen gel-embedded culture / Hepatocyte / Prostate epithelial cell / External fertilized ovem / Tyrosine aminotransferase / Suprachiasmatic nuclers / Circadian rhythm / Alginate-coated beads
Research Abstract

In this study, we developed the three-dimensional cell culture system for various types of cells using collagen gel matrix, and found that this culture system was better to maintain their physiological functions than monolayer culture system as shown by the following results.
1. To examine whether androgens directly stimulate the growth of prostate epithelial cells, a serum-free culture system was developed. The isolated mouse prostate epithelial cells were embedded in a collagen gel matrix and cultured a serum-free medium. Under these culture conditions fibroblastic growth was rare. Addition of EGF to this basal medium stimulated the growth of the epithelial cells. Further addition of dihydrotestosterone or testosterone further stimulated growth ; whereas under the monolayer cell culture conditions, androgens did not stimulate the epithelial growth.
2. The rat hepatocytes embedded in collagen gel matrix and in alginate-coated collagen gel beads maintained the secretion of albumin during … More long term period, and showed the elevation of liver specific enzyme activity including tyrosine aminotransferase. These phenomena were not observed in monolayer culture system. On the other hand, the abilities of DNA synthesis in collagen gel-embedded culture was similar to monolayer culture system, and showed the cell density-dependent inhibition of growth.
3. To grow nerve cells in suspension culture we have developed an alginate-coated collagen gel fiber culture method. Nerve cells from cfdck embryo dorsal root ganglia grew along the longitudinal axis of the fiber when the fiber width was narrow.
4. To establish the in vitro system of circadian pacemaker model in mammal, we applied the long term cell culture system of dissociated suprachiasmatic nucleus(SCN)neurons. Dispersed cells obtained from the SCN of 2 to 6 days Sprague-Dawley rats were plated on collagen-coated plastic dishes. Within 4 days after plating, cells formed three dimensional network structures and neural fibers were spread to connect each structure. The arginine vasopressin concentration in all of cultured dishes showed a clear free-running circadian rhythm with 23.5 hr mean period. While the dispersed S CN cells in collagen gel-embedded culture did not form such a network structure and showed low vasopressin secretion without circadian rhythmicity.
5. It is well known that the extremally fertilized mouse ova or one-cell-stage fertilized ova from mouse oviduct cannot continue development and that the majority of these ova stop at 2-cell-stage(2-cell block). To establish the extemal culture method for full development of fertilized ova, we applied various culture systems. The 93% of onecellrstage ovum developed to blastula stage when they were cultured inside the resected oviduct, ind the 46% of ova developed to the same stage on the incised oviduct. The only 13% of ova developed to blastula stage when they were cultured alone on collagen gel matrix, or in parallel culture widi oviduct.
From these results, it is concluded that the collagen gel-embedded culture provides good environment for various epithelial cells. To establish the culture conditions for SCN neuron and fertilized ovum, further examinations are needed. Less

  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Sakai,S.,Suzuki,M and Kohmoto,K.: "Thermodynamic analysis of the interaction of prolactin with its receptor in the rabbit mammary-gland microsomes." Biochem.J.269. 647-650 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Deng,C.Y.and Kohmoto,K.: "セルロ-スアセテ-ト膜電気泳動によるPhi,6Pgd遺伝子の同時判定と豚ハロセン遺伝子型の推定" 日蓄会報. 61. 952-955 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 渡辺 和人,榎並 淳平,長谷川 薫,山岡 貞夫: "コラ-ゲンゲル培養法の応用:肝実質細胞のコラ-ゲンゲル内包埋培養によるTAT活性の誘導" 組織培養. 16. 332-335 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Choi,T.S,Aoki,F.,Mori,M.,Yamashita,M.,Nagasawa,Y.and Kohmoto,k.: "Activation of p34^<cdc2> protein kinase activity in meiotic and mitotic cell cycles in mouse oocytes and embryos." Development. 113. 789-795 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大竹 英樹,長谷川 薫,渡辺 和人: "肝細胞における増殖関連遺伝子のクロ-ニング" 蛋白質核酸酵素. 36. 1101-1106 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki,M.,Kohmoto,K.and Sakai,S.: "Radioreceptor assay of serum prolaction using nitrocellulose membrance-immobillized mammary prolaction receptor." Anal.Biochem.200. 42-46 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sorimachi,K.,Watanabe,K.,Yamazaki,S.and Yasumura,Y.: "Inhibition of fibroblast growth by polyanions;Effect of dextran sulfate and lignin derivatives." Cell Biol.Int.Rep.16. 63-71 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murakami,H.,Kohmoto,K.and Sakai,S.: "Immunological hetero-geneity of prolactin receptors in tissues." Endocrinol.Jpn.(1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Enami,J.,Arai,O.,Saitoh,N.and Yamaoka,S.: "Growth of nerve cells cultured in alginate-coated collagen fibers." Jpn.J.Physiol.40. S30 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] watanabe,K.and Yamaoka,S.: "Circadian rhythms of vasopressin secretion in cultures of rat suprachiasmatic nucleus" Jpn.J.Physiol.40. S136 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山岡 貞夫,渡辺 和人: "視交叉上核のバゾプレッシン分泌リズム" 日本生気象学雑誌. 27(Suppl). 62 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Watanabe,K.and Yamaoka,S.: "Growth and liver-specific functions in primary culture of adult rat hepatocytes embedded in collagen gels." Jpn.J.Physiol. 41. S71 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Watanabe,K.,Koibuchi,N.and Yamaoka,S.: "Circadian rhythm of vasopressin secretion in primary cultures of rat suprachiasmatic nucleus" Jpn.J.Physiol. (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 榎並 淳平: "細胞外マトリックスの動的機能(細胞工学別冊6,板倉照〓編)乳腺分化における細胞外マトリックス" 秀潤社, 81-88 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 榎並 淳平: "新生化学講座第18巻細胞培養技術(日本生化学会編)乳腺細胞" 東京化学同人, 161-167 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 榎並 淳平: "細胞とバイオサイエンスI (日本組織培養学会編)培養細胞系における各種細胞の機能と分化:乳腺" 朝倉書店, 197-203 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] ENAMI, J.: "Effect of extracellular matrix on diferentiation of mammary gland. In "Dynamic Function of Extracellular Matrix" ed. T. Sakakura Shuujunsha Tokyo (in Japanese)" Cell Technology. Suppl. 6. 81-88 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sakai, S., Suzuki, M. and Kohmoto, K.: "Thermodynamic analysis of the interaction of prolactin with its receptor in the rabbit mammary-gland microsomes" Biochem. J.269. 647-650 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, K., Enami, J., Hasegawa, K. and Yamaoka, S.: "Primary culture of adult rat hepatocytes embedded in collagen gels -Induction of tyrosine aminotransferase. (in Japanese)" The Tissue Culture. 16. 332-335 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Choi, T. S., Aoki, F., Mori, M., Nagasawa, Y. and Kohmoto, K.: "Activation of p34^<cdc2> protein kinase activity in meiotic and mitotic cell cycles in mouse oocytes and embryos" Development. 113. 789-795 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, M., Kohmoto, K. and Sakai, S.: "Radioreceptor assay of serum prolactin using nitrocellulose membrane-immobillised mammary prolactin receptor" Anal. Biochem. 200. 42-46 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sorimachi, K., Watanabe, K., Yamazaki, S. and Yasumura, Y.: "Inhibition of fibroblast growth by polyanions ; Effect of dextran sulfate and liginin derivatives" Cell Biol. Int. Rep.16. 63-71 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Enami, J., Arai, O., Saitoh, N. and Yamaoka, S.: "Growth of nerve cells cultured in alginate-coated collagen fibers" Jpn. J. Physiol. 40. S30 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, K., and Yamaoka, S.: "Circadian rhythm of vasopressin secretion in cultures of rat suprachiasmatic nucleus" Jpn. J. Physiol. 40. S136 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamaoka, S. and Watanabe, K.: "The rhythm of vasopression in suprachiasmatic nucleus" Jpn. J. Biometeor. 27(Suppl.). 62 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, K. and Yamaoka, S.: "Growth and liver-specific function in primary culture of adult rat hepatocytes embedded in collagen gels" Jpn. J. Physiol. 41. S71 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Aoki, F., Choi, T. S., Yamashita, M. and Kohmoto, K.: "A deficiency in the mechanism for p34^<cdc2> proteinkinase activation in mouse embryo arrested in 2-cell stage" Symporium of preimplantation embryo development Aug. 15-18, 1991, Boston I-5. 27

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Choi, T. S., Aoki, F. and Kohmoto, K.: "p34^<cdc2> protein kinase in mouse oocytes and embryos" Symposium on preimplantation embryo development Aug. 15-18, 1991, Boston I-11. 30

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sobajima, T., Aoki, F. and Kohmoto, K.: "Activation of MAP kinase and H1-kinase during oocyte matulation and embryo development" Symposium on preimplantation embryo development Aug. 15-18, 1991, Boston, I-45. 47

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, K., Koibuchi, N. and Yamaoka, S.: "Circadian rhythm of vasopression in primary cultures of rat suprachiasmatic nucleus" Jpn. J. Physiolgy. 42. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, K., Yamaoka, S.: "Circadian rhythms of vasopressin secretion in long term cell cultures of rat suprachiasmatic nucleus" 9th International Congress of Endocrinology Aug. 31-Sept. 5 1992, Nice.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Enami, J.: Asakura Shoten, Tokyo. Mammary Gland. In "Cell and Bioscience" ed. The Japanese Tissue Culture Association (in Japanese), (1991)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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