PMID- 24342703 OWN - NLM STAT- MEDLINE DCOM- 20140910 LR - 20161125 IS - 1876-7753 (Electronic) IS - 1873-5061 (Linking) VI - 12 IP - 1 DP - 2014 Jan TI - Induction of dental epithelial cell differentiation marker gene expression in non-odontogenic human keratinocytes by transfection with thymosin beta 4. PG - 309-22 LID - S1873-5061(13)00168-2 [pii] LID - 10.1016/j.scr.2013.11.006 [doi] AB - Previous studies have shown that the recombination of cells liberated from developing tooth germs develop into teeth. However, it is difficult to use human developing tooth germ as a source of cells because of ethical issues. Previous studies have reported that thymosin beta 4 (Tmsb4x) is closely related to the initiation and development of the tooth germ. We herein attempted to establish odontogenic epithelial cells from non-odontogenic HaCaT cells by transfection with TMSB4X. TMSB4X-transfected cells formed nodules that were positive for Alizarin-red S (ALZ) and von Kossa staining (calcium phosphate deposits) when cultured in calcification-inducing medium. Three selected clones showing larger amounts of calcium deposits than the other clones, expressed PITX2, Cytokeratin 14, and Sonic Hedgehog. The upregulation of odontogenesis-related genes, such as runt-related transcription factor 2 (RUNX2), Amelogenin (AMELX), Ameloblastin (AMBN) and Enamelin (ENAM) was also detected. These proteins were immunohistochemically observed in nodules positive for the ALZ and von Kossa staining. RUNX2-positive selected TMSB4X-transfected cells implanted into the dorsal subcutaneous tissue of nude mice formed matrix deposits. Immunohistochemically, AMELX, AMBN and ENAM were observed in the matrix deposits. This study demonstrated the possibility of induction of dental epithelial cell differentiation marker gene expression in non-odontogenic HaCaT cells by TMSB4X. CI - Copyright (c) 2013. Published by Elsevier B.V. FAU - Kiyoshima, Tamotsu AU - Kiyoshima T AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Fujiwara, Hiroaki AU - Fujiwara H AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Nagata, Kengo AU - Nagata K AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Wada, Hiroko AU - Wada H AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Ookuma, Yukiko F AU - Ookuma YF AD - Section of Pediatric Dentistry, Division of Oral Health, Growth and Development, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Shiotsuka, Maho AU - Shiotsuka M AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan; Section of Orthodontics and Dentofacial Orthopedics, Division of Oral Health, Growth and Development, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Kihara, Makiko AU - Kihara M AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan; Section of Orthodontics and Dentofacial Orthopedics, Division of Oral Health, Growth and Development, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Hasegawa, Kana AU - Hasegawa K AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan; Department of Endodontology and Operative Dentistry, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Someya, Hirotaka AU - Someya H AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan; Section of Implant and Rehabilitative Dentistry, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. FAU - Sakai, Hidetaka AU - Sakai H AD - Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. Electronic address: hsakaiop@dent.kyushu-u.ac.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20131116 PL - England TA - Stem Cell Res JT - Stem cell research JID - 101316957 RN - 0 (Biomarkers) RN - 549LM7U24W (thymosin beta(4)) RN - 61512-21-8 (Thymosin) SB - IM MH - Animals MH - Biomarkers/metabolism MH - Calcification, Physiologic MH - *Cell Differentiation MH - Cell Line MH - *Gene Expression Regulation MH - Humans MH - Keratinocytes/*cytology/metabolism/transplantation MH - Mice MH - Mice, Nude MH - Odontogenesis/*genetics MH - RNA Interference MH - Thymosin/antagonists & inhibitors/*genetics/*metabolism MH - Tooth/cytology/metabolism MH - Transfection EDAT- 2013/12/18 06:00 MHDA- 2014/09/11 06:00 CRDT- 2013/12/18 06:00 PHST- 2013/04/04 00:00 [received] PHST- 2013/11/06 00:00 [revised] PHST- 2013/11/08 00:00 [accepted] PHST- 2013/12/18 06:00 [entrez] PHST- 2013/12/18 06:00 [pubmed] PHST- 2014/09/11 06:00 [medline] AID - S1873-5061(13)00168-2 [pii] AID - 10.1016/j.scr.2013.11.006 [doi] PST - ppublish SO - Stem Cell Res. 2014 Jan;12(1):309-22. doi: 10.1016/j.scr.2013.11.006. Epub 2013 Nov 16.