In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature.
In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature.
复制标题
牙周膜细胞在四种介质中室温保存后的体外活力、有丝分裂能力和克隆形成能力。
DOI：
10.1034/j.1600-9657.2000.016002063.x
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
H. Sarnat
中科院分区：
文献类型：
--
作者：
M. Ashkenazi;M. Marouni;H. Sarnat
外伤性撕脱牙保存材料的选择对再植成功至关重要。本研究的目的是比较四种推荐的储存介质（Hank平衡盐溶液[HBSS]、培养基、α最小必需培养基[α-MEM]和ViaSpan）在室温（22 ℃）下保存培养的牙周膜成纤维细胞（PDLF）的有效性。PDLF从提取的健康人牙齿的外植体获得。将具有汇合的PDLF的板在室温下在各种培养基中浸泡2、8和24小时。对照组在37 ℃下与培养基一起孵育。孵育后，通过台盼蓝排除试验测定细胞的活力。然后分析活细胞的促有丝分裂（用胸苷）和克隆形成能力（通过培养一个细胞/孔）。储存长达24 h的PDLF的活力在所有测试的培养基中相当，并且差异限于1%-3%。在各种培养基中储存长达24小时的PDLF的有丝分裂原性与对照组在统计学上相当。储存8小时后，差异限于2%-9%，除了α-MEM组的促有丝分裂能力比对照组低23%-29%。将储存时间延长至24 h，细胞的促有丝分裂性进一步降低22%-47%。PDLF在不同培养基中保存2-8 h后，其克隆形成能力与对照组相当。但24小时后，细胞的克隆形成能力下降了14%-66%。在促有丝分裂和克隆形成能力中观察到类似的降低趋势，尽管仅在克隆形成能力中具有统计学显著性。培养基和ViaSpan，其次是HBSS，是最有效的保存PDLF的克隆形成能力后，24小时的存储。保存24 h后，α-MEM组的克隆形成能力最低（66%，P < 0.0025）。总之，培养基，其次是HBSS和ViaSpan，是保存PDLF在室温下储存长达24 h的活力、促有丝分裂性和克隆形成能力的最有效培养基。在储存于α-MEM中的PDLF中发现最低的功能能力。
The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to compare the effectiveness of four recommended storage media (Hank's balanced salt solution [HBSS], culture medium, alpha minimal essential medium [alpha-MEM], and ViaSpan) to preserve cultured periodontal ligament fibroblasts (PDLF) at room temperature (22 degrees C). PDLF were obtained from explants of extracted healthy human teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at room temperature. A control group was incubated with culture medium at 37 degrees C. After incubation, viability of the cells was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Viability of PDLF stored up to 24 h was comparable in all tested media, and the differences were limited to 1%-3%. PDLF stored for up to 24 h in various media had statistically comparable mitogenicity to the control group. After 8 h of storage, the differences were limited to 2%-9%, except for the alpha-MEM group which had 23%-29% lower mitogenic capacity compared to the control group. Increasing the storage time up to 24 h further decreased the mitogenicity of the cells by 22%-47%. The highest mitogenicity after 24 h of storage was found in PDLF stored in culture medium or HBSS, and the lowest in alpha-MEM. PDLF stored for 2-8 h in various media had a comparable clonogenic capacity to the control group. However, after 24 h, the cells' clonogenic ability dropped by 14%-66%. A similar trend of reduction was noted in the mitogenic and clonogenic capacity, although it was statistically significant only in the clonogenic capacity. Culture medium and ViaSpan, followed by HBSS, were the most effective in preserving the clonogenic capacity of PDLF after 24 h of storage. The lowest clonogenic capacity after 24 h of storage was in the alpha-MEM group (66%, P < 0.0025). In conclusion, culture medium, followed by HBSS and ViaSpan, was the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for up to 24 h at room temperature. The lowest functional abilities were found in PDLF stored in alpha-MEM.