Development of polymer medicines production system ulilzing polarized epithelial culturing method.

1993 Fiscal Year Final Research Report Summary

• Project/Area Number: 04556009
• Research Category: Grant-in-Aid for Developmental Scientific Research (B)
• Allocation Type: Single-year Grants
• Research Field: 応用生物化学・栄養化学
• Research Institution: Nagoya University
• Principal Investigator: KITAGAWA Yasuo Nagoya University Bioscience Center, Professor, 生物分子応答研究センター, 教授 (50101168)
• Co-Investigator(Kenkyū-buntansha): MIKI Kiyoshi Nagoya University Bioscience Center, Research Associate, 生物分子応答研究センター, 助手 (30212228) ; UEDA Masatugu Snow-Brand Milk Co.Ltd.Director, 生物科学研究所, 主査
• Project Period (FY): 1992 – 1993
• Keywords: Erythropoietin / Polarized secretion / Polymer medicine / Glycen-processing / 糖鎖修飾

Research Abstract

Polarized epithelial cells are characterized by two plasma membrane domains, apical and basolateral domains, having different structure and functions. The polarity of epithelial cells is not only restricted to plasma membrane components but secretory proteins are also released in a polarized fashion. MDCK cells are a typical epithelial cell line derived from dog kidney and have been widely used to investigate molecular mechanism of vectorial transport of membrane lipids and proteins. Erythropoietin (Epo) is a major physiological regulator of erythropoiesis by stimulating proliferation and differentiation of erythroid precursor cells. We here found that recombinant human Epo is preferentially secreted from the apical domain of MDCK cells. Taking advantage of this polarized secretion, we have developed a pilot system of producing Epo in industrial scale. We have collected basic information of deigning hollow-fiber system in which Epo will be released either inside or outside of the fiber.

Research Products

• [Publications] Taniguchi.Y.,Miki.K.,and Kitagawa.Y.: "Transient gene expression by SV40 promoter characterizes sequential differentiation of embryonic carcinoma F4 into primitive and viscrral endoderm." Experimental Cell Research. 204. 286-294 (1993)
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• [Publications] Taniguchi.Y.and Kitagawa.Y.: "The differentiation-dependent inhibition of SV40 promoter/enhancer aofivity in 3T3-L1 cells is mediated through promoter and not enhancer sequence" Cytotechnology. 11. 175-182 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 北川泰雄,佐野祐子,岡野正樹,上田正次,佐々木隆生: "細胞のうらおもてとエリスロポエチン生産" 化学と生物. 31. 205-212 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Jeon.H.,Ono.M.,Kumagai.C.,and Kitagawa.Y.: "Subunit assembly of laminin subunits in cultured animal cells." Animal Cell Technology;Basic & Applied Aspects. 5. 41-48 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Inagaki.T.,Matsushima M.,Ohshima.M.,and Kitagawa.Y.: "Regulation of mitochordrial gene oxpression by mterforouos" Animal Cell Technology;Basic & Applied Aspects. 5. 49-55 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ono.M.,and Kitagawa.Y.: "Dextrou sulfate accelerates adipose conversion of 3T3-L1 Cells." Bioscience,Biotechnology and Biochemistry. (印刷中).
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kitagawa, Y., Sano, Y., Ueda, M., Higashio, K., Narita, H., Okano, M., Matsumoto, S.and Sasaki, R.: "N-Glycosylation of erythropoietin for its secretion from apical domain of Madin-Darby canine kidney cells." Experimental Cell Research. (in press). (1994)
  • Description: 「研究成果報告書概要(欧文)」より