신생 백서 척수후근절의 슈반세포 배양을 위한 Ara-C 분열억제제의최적 효과에 대한 연구
The most appropriate antimitotic treatment of Ara-C in schwann cell-enriched culture from dorsal root ganglia of new born rat
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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%±8.09% in P4 group to 65.5%±24.08% in P2 group), compared with that obtained in the abscence of Ara-C (17.6%±6.03%) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to 56.22%±0.67% and GFAP positive cells to 66.46%±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.
참고문헌 (25)
- "Schwann cells as regulators of nerve development", 96, pp.17~24 (2002)
- "Making of a Schwann", 12, pp.84~86 (1996)
- "Effect of Schwann cells in the enhancement of peripheral-nerve regeneration", 12, pp.439~446 (1996)
- "The influence of heregulins on human Schwann cell proliferation", 15, pp.1329~1340 (1995)
- "Studies on culture rat Schwann cells Establishment of purified populations from cultures of peripheral nerve", pp.105~118 (1979)
- "Purification and expansion of human Schwann cells in vitro", 1, pp.80~83 (1995)
- "Selective expansion and long-term culture of human Schwann cell from sural nerve biopsies", 38, pp.674~678 (1995)
- "Axolemma is a mitogen for human Schwann cells", pp.449~452
- "Schwann cell-enriched cultures from adult human peripheral nerve a technique combining short enzymatic dissociation and treatment with cytosine arabinoside", 114, pp.1~8 (2002)
- "Isolation of activated adult Schwann cells and a spontaneously immortal Schwann cell clone", pp.231~238 (1992)
- "Improved protocol for schwann cell isolation and proliferation from rat dorsal root ganglia in vitro", 25, pp.265~272 (2003)
- "Culture of animal cells a manual of basic technique", pp.9~18 (2000), Wiley-Liss Inc
- "Fabrication and implantation of gel-filled nerve guidance channels Methods in Molecular Medicine", 18 (1999), Humana Press Inc
- "Functional recovery following nerve injury and repair by silicon tubulization: comparison of laminin-fibronectin, dialyzed plasma, collagen gel, and phosphate buffered solution", 26, pp.117~122 (1999)
- "Two-dimensional versus three-dimensional cell counting:a practical perspective", 24, pp.11~17 (2001)
- "Three-dimensional counting:an accurate and direct method to estimate numbers of cells in sectioned material", 278, pp.344~352 (1988)
- "Density dependent regulation of human Schwann cell proliferation", 30, pp.165~177 (2000)
- "Improved method for harvesting human Schwann cells from mature peripheral nerve and expansion in vitro", 17, pp.327~338 (1996)
- "Successfully culturing Schwann cells from adult peripheral nerve", 30, pp.15~19 (1998)
- "Characterization of the technique involved in isolating Schwann cells from adult human peripheral nerve", 68, pp.21~26 (1996)
- "Viability of cultured nerve grafts an assessment of proliferation of Schwann cells and fibroblasts", 19, pp.356~363 (1999)
- "Influence of collagen and laminin gels concentration on nerve regeneration after resection and tube repair", 149, pp.243~252 (1998)
- "A Schwann cell-seeded intrinsic framework and its satisfactory biocompatibility for a bioartificial nerve graft", 21, pp.6~11 (2001)
- "Isolation and serum-free culture of primary Schwann cells from human fetal peripheral nerve", 158, pp.1~8 (1999)
- "In vivo predegeneration of peripheral nerves an effective technique to obtain activated Schwann cells for nerve conduits", 89, pp.17~24 (1999)