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Development of a gene transfer method in utero and a culture method for reconstructed hair germs for study of the process of hair development

Research Project

Project/Area Number 13670887
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Dermatology
Research InstitutionShimane University

Principal Investigator

MATSUZAKI Takashi  Shimane Univ., Fac. Life & Environmental Science, Assistant prof., 生物資源科学部, 講師 (90241249)

Co-Investigator(Kenkyū-buntansha) IHARA Setsunosuke  Shimane Univ., Fac. Life & Environmental Science, Professor, 生物資源科学部, 教授 (90101295)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
Keywordsretrovirus / gene transfer in utero / keratinocyte / hair follicle / cell differentiation / reconstraction
Research Abstract

In order to establish a convenient method for gene transfer into the embryonic ectoderm, retroviruses containing a reporter gene, Lac-Z or GFP, were injected into the amniotic cavity of mice at 8.0 - 13.5 days of gestation. Improvement of gene transfer efficiency by retroviruses was observed in the limb, tail, and back skin when applied with poly-L-lysine. Adding poly-L-ornithine was also effective, but only in the tail skin. Although efficiency of gene transfer enhanced as virus concentration increased up to 100,000 cfu, it reduced if higher number of virus was injected. For X-gal reaction, optimal duration of fixation with 2% PFA was 3.5 hrs for a whole embryo at 16.0 days of gestation. LacZ-positive cells were detected in the skin with various patterns, which suggests that origin of periderm was distinct from one of epidermis. Both lacZ- and GFP-positive cells were observed even in the skin of 11-weeks-old mice. As they were also detected in the hair follicles, retrovirus infection in utero is thought to be an excellent method to introduce exogenous genes into the embryonic ectoderm and to trace cell lineages in the hair follicles.
In the experiments of two-step culture, it became clear that the most important point of culture was milder but complete dissociation of hair germ cells. This condition was achieved by treating the hair germ-containing skin with 0.25% trypsin at 4℃ for six hrs. Dissociated hair germ cells could self-assemble and make an aggregate in the reconstructed skin after rotation culture, which was clearly demonstrated by mixing GFP-tagged hair germ cells. However, we failed to develop hair follicles in them during floatation culture. Thus, it is thought to be important to keep an adequate interaction between self-assembled hair germ and hair-inducers such as dermal papillae.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Kiyokazu Morioka: "Localization of cathepsins B, D L, LAMP-1 and m-calpain in developing hair follicles"Acta Histochem.Cytochem.. 34. 337-347 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "新規発毛剤スペラゲン707のマウス休止期毛包に対する発毛促進効果"島根大学生物資源科学部研究報告. 6. 17-22 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "ヘアーバイオロジーからみた毛髪(1)"毛髪科学. 89. 8-15 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "ヘアーバイオロジーからみた毛髪(2)"毛髪科学. 90. 3-11 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "ヘアーバイオロジーからみた毛髪(3)"毛髪科学. 91. 3-10 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "毛髪の生物学"フレグランスジャーナル. 30(8). 11-15 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K,Morioka., K,Sato-Kusubata., S,Kawashima., T,Ueno., E,Kominami., H,Sakuraba., S,Ihara.: "Localization of cathepsins B, D L, LAMP-1 and m-calpain in developing hair follicles."Acta Histochem. Cytocham. 34. 337-347 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T,Matsuzaki.: "Effects of a novel hair restorer Spelagen 707 on hair growth initiation to telogen pelege follicles in mice"Bull. Fac. Life. Environ. Sci. Shimane Univ. (Jpn). 6. 17-22 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T,Matsuzaki.: "Hair dissected from molecular biology (1)"Hair Science, (Jpn). 89. 8-15 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T,Matsuzaki.: "Hair dissected from molecular biology (2)"Hair Science, (Jpn). 90. 3-11 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T,Matsuzaki.: "Hair dissected from molecular biology (3)"Hair Science, (Jpn). 91. 3-10 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T,Matsuzaki.: "Hair biology"Fragrance J., (Jpn). 30(8). 11-15 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 松崎 貴: "ヘアーバイオロジーから見た毛髪(2)"毛髪科学. 90. 3-11 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 松崎 貴: "ヘアーバイオロジーから見た毛髪(3)"毛髪科学. 91. 3-10 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 松崎 貴: "毛髪の生物学"Fragrance J.. 30・8. 11-15 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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