title: Scaling-up of dental pulp stem cells isolated from multiple niches.
abstract: 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".
authors: Laboratory of Genetics, Butantan Institute, Sao Paulo, SP, Brazil. nelson_lizier@yahoo.com.br
authors: Nelson F
authors: NF
authors: Lizier
authors: Alexandre
authors: A
authors: Kerkis
authors: Cícera M
authors: CM
authors: Gomes
authors: Josimeri
authors: J
authors: Hebling
authors: Camila F
authors: CF
authors: Oliveira
authors: Arnold I
authors: AI
authors: Caplan
authors: Irina
authors: I
authors: Kerkis
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002648:Child; D020219:Chondrogenesis / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D003782:Dental Pulp / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D024510:Muscle Development / Q000187:drug effects; D055153:Stem Cell Niche / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors
chemical_list: D015415:Biomarkers; D003470:Culture Media; D001973:Bromodeoxyuridine