Introduction:Dental pulp stem cells from humans possess self-renewal and versatile differentiation abilities. These cells, known as DPSC, are promising for tissue engineering due to their outstanding biological characteristics and ease of access without significant donor site trauma. Existing methods for isolating DPSC mainly include enzyme digestion and explant techniques. Compared with the enzymatic digestion technique, the outgrowth method is less prone to cell damage and loss during the operation, which is essential for DPSC with fewer tissue sources.Methods:In order to maximize the amount of stem cells harvested while reducing the cost of DPSC culture, the feasibility of the optimized explant technique was evaluated in this experiment. Cell morphology, minimum cell emergence time, the total amount of cells harvested, cell survival, and proliferative and differentiation capacity of DPSC obtained with different numbers of explant attachments (A1-A5) were evaluated.Results:There was a reduction in the survival rate of the cells in groups A2-A5, and the amount of harvested DPSC decreased in A3-A5 groups, but the DPSC harvested in groups A1-A4 had similar proliferative and differentiation abilities. However, starting from group A5, the survival rate, proliferation and differentiation ability of DPSC decreased significantly, and the adipogenic trend of the cells became more apparent, indicating that the cells had begun to enter the senescence state.Discussion:The results of our study demonstrated that the DPSC obtained by the optimized explant method up to 4 times had reliable biological properties and is available for tissue engineering.

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如何充分利用牙髓干细胞：基于外植体技术的优化细胞培养方法

简介：来自人类的牙髓干细胞具有自我更新和多种分化能力。这些被称为 DPSC 的细胞由于其出色的生物学特性并且易于获取而不会造成严重的供体部位创伤，因此在组织工程方面具有广阔的前景。现有分离DPSC的方法主要包括酶消化和外植体技术。与酶消化技术相比，生长法在操作过程中不易发生细胞损伤和丢失，这对于组织来源较少的DPSC至关重要。 方法：为了最大限度地提高干细胞的收获量，同时降低DPSC的成本培养，本实验评估了优化的外植体技术的可行性。评估不同数量的外植体附着体（A1-A5）获得的DPSC的细胞形态、最短细胞出现时间、收获的细胞总量、细胞存活率以及增殖和分化能力。结果：存活率降低A2-A5组细胞数量减少，A3-A5组收获的DPSC量减少，但A1-A4组收获的DPSC具有相似的增殖和分化能力。但从A5组开始，DPSC的存活率、增殖和分化能力明显下降，细胞成脂趋势更加明显，表明细胞已开始进入衰老状态。 讨论：我们的研究结果证明通过优化外植体方法高达4次获得的DPSC具有可靠的生物学特性，可用于组织工程。