Search

Made available in DSpace on 2013-08-28T14:12:40Z (GMT). No. of bitstreams: 1 WOS000305892100123.pdf: 1025211 bytes, checksum: f3114ad3d761d448ea4009979a8c8d68 (MD5) Made available in DSpace on 2013-09-30T18:31:32Z (GMT). No. of bitstreams: 2 WOS000305892100123.pdf: 1025211 bytes, checksum: f3114ad3d761d448ea4009979a8c8d68 (MD5) WOS000305892100123.pdf.txt: 56945 bytes, checksum: e71c3f4e9d87411582be429d4ea1669f (MD5) Previous issue date: 2012-06-29 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:44:59Z No. of bitstreams: 2 WOS000305892100123.pdf: 1025211 bytes, checksum: f3114ad3d761d448ea4009979a8c8d68 (MD5) WOS000305892100123.pdf.txt: 56945 bytes, checksum: e71c3f4e9d87411582be429d4ea1669f (MD5) Made available in DSpace on 2014-05-20T13:44:59Z (GMT). No. of bitstreams: 2 WOS000305892100123.pdf: 1025211 bytes, checksum: f3114ad3d761d448ea4009979a8c8d68 (MD5) WOS000305892100123.pdf.txt: 56945 bytes, checksum: e71c3f4e9d87411582be429d4ea1669f (MD5) Previous issue date: 2012-06-29 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Dental pulp (DP) can be extracted from child's primary teeth (deciduous), whose loss occurs spontaneously by about 5 to 12 years. Thus, DP presents an easy accessible source of stem cells without ethical concerns. Substantial quantities of stem cells of an excellent quality and at early (2-5) passages are necessary for clinical use, which currently is a problem for use of adult stem cells. Herein, DPs were cultured generating stem cells at least during six months through multiple mechanical transfers into a new culture dish every 3-4 days. We compared stem cells isolated from the same DP before (early population, EP) and six months after several mechanical transfers (late population, LP). No changes, in both EP and LP, were observed in morphology, expression of stem cells markers (nestin, vimentin, fibronectin, SH2, SH3 and Oct3/4), chondrogenic and myogenic differentiation potential, even after cryopreservation. Six hours after DP extraction and in vitro plating, rare 5-bromo-2'-deoxyuridine (BrdU) positive cells were observed in pulp central part. After 72 hours, BrdU positive cells increased in number and were found in DP periphery, thus originating a multicellular population of stem cells of high purity. Multiple stem cell niches were identified in different zones of DP, because abundant expression of nestin, vimentin and Oct3/4 proteins was observed, while STRO-1 protein localization was restricted to perivascular niche. Our finding is of importance for the future of stem cell therapies, providing scaling-up of stem cells at early passages with minimum risk of losing their stemness. Instituto Butantan, Genet Lab, São Paulo, Brazil Fed Univ São Paulo UNIFESP, Dept Morphol, São Paulo, Brazil State Univ São Paulo UNESP, Dept Orthodont & Pediat Dent, Araraquara, SP, Brazil Case Western Reserve Univ, Dept Biol, Skeletal Res Ctr, Cleveland, OH 44106 USA State Univ São Paulo UNESP, Dept Orthodont & Pediat Dent, Araraquara, SP, Brazil FAPESP: 10/51051-6