Journal of the Korean Association of Oral and Maxillofacial Surgeons

The Korean Association of Oral and Maxillofacial Surgeons (대한구강악안면외과학회)
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THE MOST APPROPRIATE ANTIMITOTIC TREATMENT OF ARA-C IN SCHWANN CELL-ENRICHED CULTURE FROM DORSAL ROOT GANGLIA OF NEW BORN RAT

신생 백서 척수후근절의 슈반세포 배양을 위한 Ara-C 분열억제제의 최적 효과에 대한 연구

  • Kim, Soung-Min (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Lee, Jong-Ho (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Ahn, Kang-Min (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Kim, Nam-Yeol (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Sung, Mi-Ae (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Hwang, Soon-Jeong (Department of Oral & Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Kim, Ji-Hyuck (Department of Oral & Maxillofacial Surgery, College of Dentistry, Kangnung National University) ;
  • Jahng, Jeong-Won (Department of Pharmacology, Yonsei Medical School)
  • 김성민 (서울대학교 치과대학 구강악안면외과) ;
  • 이종호 (서울대학교 치과대학 구강악안면외과) ;
  • 안강민 (서울대학교 치과대학 구강악안면외과) ;
  • 김남열 (서울대학교 치과대학 구강악안면외과) ;
  • 성미애 (서울대학교 치과대학 구강악안면외과) ;
  • 황순정 (서울대학교 치과대학 구강악안면외과) ;
  • 김지혁 (강릉대학교 치과대학 구강악안면외과) ;
  • 장정원 (연세대학교 의과대학 약리학교실)
  • Published : 2004.04.30
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Abstract

Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

Keywords

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