Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.
Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.
复制标题
牙齿组织工程：血液制品作为人牙髓干细胞组织培养基补充剂的重要性。
DOI：
10.1089/ten.tea.2014.0617
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发表时间：
2015
影响因子：
--
通讯作者：
Duailibi,SilvioEduardo
中科院分区：
文献类型：
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作者：
Pisciolaro,RicardoLuiz;Duailibi,MonicaTalarico;Novo,NeilFerreira;Juliano,Yara;Pallos,Debora;Yelick,PamelaCrotty;Vacanti,JosephPhillip;Ferreira,LydiaMasako;Duailibi,SilvioEduardo
使用细胞进行组织工程 (TE) 和细胞治疗的目标之一是优化细胞培养介质。本研究比较了三种不同的血液制品补充剂，以改善牙齿 TE 应用中培养的人牙髓干细胞的细胞增殖和防止 DNA 损伤。首先通过流式细胞术将来自牙髓的人类细胞定性为成体干细胞（外间充质混合来源）。接下来，测试了四种不同的细胞培养条件：I，无补充剂； II、补充胎牛血清； III、同种异体人血清； IV，自体人血清。然后对培养细胞的细胞增殖、矿化结节形成和集落形成单位 (CFU) 能力进行表征。培养 28 天后，进行彗星测定以评估细胞 DNA 可能的损伤。我们的结果表明，方案 IV 比方案 I 实现了更高的细胞增殖 (p= 0.0112)。方案II和III导致比方案I更高的细胞增殖，但相对于方案IV没有发现统计差异。彗星测定显示，与方案 II 和 III 相比，使用方案 IV 培养的细胞损伤较少。在协议 II 上观察到的损坏百分比明显高于所有其他协议。分别使用方案 IV (p= 0.0018) 和 III 时 CFU 能力最高，并且与方案 II 和 III 相比，使用方案 IV 观察到最高的矿化程度。方案IV导致细胞增殖显着改善，并且没有观察到细胞损伤。这些结果表明，人类血液制品补充剂可以用作培养成人牙齿干细胞的可行补充剂。
One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I (p= 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV (p= 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.