Browse > Article

Establishment of Immotalized Human Gingival Fibroblast Cell Lines  

Song, Jae-Bong (Department of Periodontology, School of Dentistry, Wonkwang University)
Kim, Hyun-A (Department of Periodontology, School of Dentistry, Wonkwang University)
Hyun, Ha-Na (Department of Periodontology, School of Dentistry, Wonkwang University)
Kim, Eun-Cheol (Department of Oral Pethology, School of Dentistry, Wonkwang University)
You, Hyung-Keun (Department of Periodontology, School of Dentistry, Wonkwang University)
Shin, Hyung-Shik (Department of Periodontology, School of Dentistry, Wonkwang University)
Publication Information
Journal of Periodontal and Implant Science / v.32, no.3, 2002 , pp. 603-614 More about this Journal
Abstract
Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamHl and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful　transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.
Keywords
Gingival Fibroblast; Cell line; Immortalization; HPV; Growth; proliferation;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Boyko GA, Melcher AH, Brunette DM. Formation of new periodontal ligament by periodontal ligament cells implanted in vivo after culture in vitro. A preliminary study of transplanted roots in the dog. J Periodontol Res 16: 73-88, 1981   DOI
2 Polson AM, Caton J. Factors influencing periodontal repair and regeneration. J Periodontol 53: 617-625, 1982   DOI
3 Ragnarsson B, Carr G, Daniel JC. Isolation and growth of human perodontal ligament cells in Vitro. J Dent Res 64: 1026-1030, 1985   DOI   ScienceOn
4 Nanba H, Nomura Y, Kinoshita M, Shimizu H, Ono K, Goto H, Aral H, Takigawa M, Murayama Y. Periodontal Tissues and Sex Hormones. The effects of Sex Hormones on Metbolism of Fibroblasts Derived from periodontal ligament. J, Japan Ass Periodont 31: 166-175, 1989   DOI
5 Nojima N, Kobayashi M, Shionome M, Takahashi N, Suda T, Hasega K. Fibroblastic cells derived from bovine periodontal ligaments have the phenotypes of osteoblasts, J Periodont Res 25: 179-185, 1990   DOI   PUBMED
6 Gottolw J, Nyman S, Karring J, Lindhe J. New attachment formation as the result of controlled tissue Regeneration. J Clin Periodontol 11: 494-503, 1984   DOI
7 Wergedal JE, Baylink DJ. Characterization of cells isolated and cultured from human bone Proc Soc Exp Biol Med 176: 60-69, 1984   DOI   PUBMED
8 Tasaka S. Establishment from mouse periodontal ligament of an immortalized cell line(MPDL-22) expressing physiological differentiation phenorype(thesis), J Osaka Univ Dent Sch 41: 118-135, 1996
9 Meanwell CA. The epidemiology of human papillomavirus infection in relation to cervical cancer. Cancer Surv 7: 481-497, 1988   PUBMED
10 Kellokoski JK, Syrjanen SM, Chang F, Yliskoski M, Syrjanen KJ. Southern blot hybridization and PCR in detection of human papilllornavirus infection in wonen with genital infection. J Oral PathoI 21:459-464, 1992   DOI   ScienceOn
11 Sjryanen S. Viral infection in oral mucosa. ScanJ Dent Res 100: 17-31, 1992
12 Hawley-Nelson P, Vousden KH, Hubbert NL, Lowy DR, Schiller JT. HPV 16 E6 and E7 proteins cooperate to immortalize human foreskin keratinocytes, EMBO J. 8: 3905-3910. 1989   PUBMED
13 Nyman S, Lindhe J, Karring T, Rylander H. New attachment following surgical treatment of human periodontal disrase. J Clin Periodontol 9: 290-296, 1982   DOI
14 Aukhil I, Simpson DM, Schaberg TV. An experimental study of new arrachment procedure in beagle dogs. J Periodont Res 18: 643-654, 1983   DOI
15 McCormick, JJ, Maher VM, Towards an under standing of the malignant transformation of the malignant transformation of diploid human fibroblasts. Mutat Res 199: 273-291, 1988   DOI   PUBMED
16 Brunette DM, Janoza RJ, Marmary Y, Chan J, Melcher AH. Interactions between epithelial and fibroblast-like cells in cultures derived from monkey periodontal ligament. J Cell Sci 27: 127-140, 1977
17 Marmary Y, Brunette DM, Heersche JNM. Differences in vitro between cells derived from periodontal ligament and skin of Macaca Irus, Archs Oral Biol 21: 709-716, 1976   DOI   ScienceOn
18 Gould TRL. Ultrastructural characteristics of progenitor cell populations in the periodontal ligament. J Dent Res 62: 873-876, 1983   DOI   PUBMED   ScienceOn
19 Piche JE, Carnes DL, Graves DT. Characteristics of a non-fibroblast cell populatin derived from human periodontal ligament. J Dent Res 66: 356(Abstr No 1998), 1987
20 Band V, De Caprio JA, Delmolino L, Kulesa V, Sager R. Loss of p53 protein in human papillomavirus type 16 E6-immortalized human mammary epithelial cells. J Virol 65: 6671-6676. 1987
21 Melcher Ah, On the repair potential of periodontal tissues. J Periodontol 47: 256-260, 1976   DOI   PUBMED
22 Isidor F, Karring T, Nyman S, Lindhe J. The significance of coronal growth of periodontal ligament tissue for new attachment formation. J Periodontol B: 145-150, 1986
23 Brunette DM, Melcher AH, Moe HK, Cultrue and origin of epithelium-like and fibroblast-like cells from porcine periodontal ligament explants and cell suspension. Archs Oral Bioi 21: 393-400, 1976   DOI   ScienceOn
24 Schwarz E, Freese UK, Gissmann L, Mayer W, Roggenbuck B, Stremlau A, Hausen H. Structure and transcription of HPV sequences in cervical cancer cells. Nature 314: 111-114, 1985   DOI   ScienceOn
25 McCulloch CAG. Progenitor cell populations in the periodontal ligament of mice. Anat Rec 211: 258-262, 1985   DOI   ScienceOn
26 Jacobsen N, Goldhaber P. Bone resorption induced by epithelial and connective tissue from human gingiva in vitro. J Dent Res 51: 1683-1689, 1972   DOI   ScienceOn
27 Naraynan AS, Nakae H, Miki Y. Biology of the gingiva : The connective tissue in health and disease and molecular aspects of periodontal regeneration and reattachment. Recent Advances in Clinical Periodontology, Elsevier Science Publishers 51-61, 1988
28 Gottlow J, Nyman S, Lindhe J, Karring T, Wennstrom J. New attachment formation in the human periodontium by guided tissue Regeneration. case report. J Clin Periodontol 13: 604-616, 1986   DOI
29 Park NH, Min BM, Li SL, Huang MZ, Cherrick HM, Doniger J. Immortalization of normal human oral keratinocyte with type 16 human papillornavirus, Cariconegensis 12: 1627-1631, 1991   DOI   ScienceOn
30 Shay JW, Wright WE, Brasiskyte D, Van Der Haegen BA. E6 of human papillomavirus in human mammary epithelial cells but in human fibroblasts. Oncogene 8: 1407-1413, 1993
31 Mariotti A, Cochran Dl. Characterization fo fibroblasts derived from human periodontal ligament and gingiva. J Periodontol 61: 103-111, 1990   DOI
32 Bryan TM, Reddek RR. SV40-induced irrunortalization of human cells. Crit Rev Oncog 5: 331-357, 1994   DOI
33 Phillips MA, Stewart BE, Qin Q, Chakrvarty R, Floyd EE, Jetten AM. Rice RH. Primary structure of keratinocyte transglutaminase. Proc Natl Acad Sci USA 87: 9333-93337, 1990   DOI   ScienceOn
34 Rose GG, Yamasak A, Pinero Gh, Mahan CJ. Human periodontal ligament cells in vitro. J Periodont Res 22: 20-28, 1987   DOI
35 Arnold LF, Baram P, In vitro culture of periodontal ligament cells. J Dent Res 51: 953-959, 1972   DOI   PUBMED   ScienceOn
36 Piche JE, Graves DT. Growth and biochemical Characteristics of cells derived from human periodontia. J Dent Res 67: 243(Abstr No. 1044), 1988
37 Piche JN, Carnes D, Land Garces DT. Initial Characterization of cells Derived from Human periodontia. J Dent Res 68: 761-767, 1989   DOI   ScienceOn
38 Fusenig NE, Boukamp KJ. Multuple stages and geneic alteration in immortalization, malignant transformation, and tumor progression of human skin keratinocyte. Mol Carcinog 23: 144-158, 1998   DOI   ScienceOn
39 Gould TRL, Melcher AH, Brunette DM. Migration and division for progenitor cell populations in periodontal ligament after wounding. J Periodont Res 15: 20-42, 1980   DOI
40 Bowers GM, Schallhorn RG, Mellonig JT. Histologic evaluation of new attachment in human intrabony defect. A literature review. J Periodontol Sf: 509-514, 1982
41 Woodworth CD, Doniger J, Dipaolo JA. Immortalization of human foreskin keratiocytes by various human papilloma virus DNA correspond to their association with cervical carcinoma. J Virol 86:159-164, 1989
42 Carlens DL, Nydegger JR, Macdougall M. Establishment of immortalized cell population from periodontla ligament . J Dent Res 76: 298, 1997
43 Freshney RI. Culture of animal cells. A manual of basic technique. In: Wiley-Liss, New York, 1987
44 Munger K, Phelps WC, Bubb V, Howley PM, Schlegel, R. The E6 and E7 genes of the human papillomavirus type 16 together are necessary and sufficient for transformation of primary human Keratinocytes, J Virol 63: 4417-4421. 1989   PUBMED
45 Aukhil I, Simpson DM, Suggs C, Pettersson E. In vivo differentiation of progenitor cells of the periodontal ligament. An experimental study using physical barriers. J Clin Periodontol 13: 862-868, 1986   DOI
46 Nosagaard H, Clark BFC, Rattan AlS. Distinction between differentiation and senecence and the abscence of increased apoptosis in human keratinocytes undergoing cellular aging in vitro. Exp Gerontology 31: 563-570, 1996   DOI   ScienceOn
47 Watanabe S, Kanda T, Yoshiike K. Human papillomavirus type 16 transformation of primary human embryonic fibroblasts requires expression of open reading frames E6 and E7. J Virol 63: 965-969. 1989   PUBMED
48 Harms W, Rothanmel T, Miller K, Harste G, Grassman M, Heim A. Chracterization of human myocradial fibroblasts immortalized by human HPV16 E6-E7 gens. Exp Cell Res 268; 252-261, 2001   DOI   ScienceOn
49 Somerman MJ, Archer SY, Imm GR, Forst RA. A comparative study of human periodontal ligament cells and gingival fibroblasts in Vitro. J Dent Res 67: 66-70, 1988   DOI   ScienceOn
50 Blomlof I, Otteskog P Composition of Human Periodontal Ligament Cells in tissue Culture. Scand, J Dent Res 89: 43-47, 1981
51 Auf'mkolk B, Hauachka PV, Schwartz ER. Characterization of human bone cells in culture. Calcif Tissue Int 37: 228-235, 1985   DOI   ScienceOn
52 Shiga T, SHirasawa H, SHimizu K, Dezawa M, Masuda Y, Simizu B. Normal Human Fibroblasts Immortalized by Introduction of Human Papillomavirus Type 16 (HPV-16) E6-E7 Genes. Micribiol ImmunoI 41; 313-319, 1997   DOI
53 Melcher AH. Repair of wounds in the periodontium of the rat. Influence of periodontal ligament on osteogenesis. Achs Oral Biol 15: 1183-1204, 1970   DOI   ScienceOn
54 Brunette DM, Heersche JNM, Purdon AD, Sadek J, Moe HK, Assuras IN. In vitro cultural parameters and protein and prostaglandin secretion of epithelial cells derived from porcine ress of malassez. Archs Oral Biol 24: 199-203, 1979   DOI   ScienceOn
55 박 준봉외 17인 : 치주과학, 지영문화사 6-59, 1988