| 张倩,汪俊.不同直径的培养皿对三维牙周膜细胞团培养的影响[J].口腔材料器械杂志,2012,21(3):140-144. |
| 不同直径的培养皿对三维牙周膜细胞团培养的影响 |
| Effect on different diameters of culture dish in three-dimensional periodontal ligament cell pellet cultivation |
| 投稿时间：2012-02-22 修订日期：2012-03-07 |
| DOI：10.11752/j.kqcl.2012.03.08 |
| 中文关键词: 培养皿直径 牙周膜细胞团 牙周再生 荧光染色 聚合酶链反应 |
| 英文关键词:Culture dish diameter PDL cell pellet Periodontal regeneration Fluorescent staining PCR |
| 基金项目:上海市重点学科建设项目(编号:S30206) |
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| 中文摘要: |
| 目的： 通过一系列方法评价不同直径的培养皿对三维牙周膜细胞团培养的影响。方法：对人牙周膜细胞进行分离,培养和鉴定。用含有维生素C的DMEM培养液分别将牙周膜细胞培养于直径为3.5cm、6cm及10cm的培养皿内,2周后,从皿底部刮取细胞层,分别放于离心管中继续培养1周,以形成密实的细胞团。然后,分别通过形态学检测、荧光染色、MTT以及PCR 4种方法来检测不同细胞团里的细胞活性和牙周再生相关基因表达。结果： 不同细胞团的活性和其它特征差别较明显,其中直径为3.5em培养皿培养出的细胞团相比另外两组有更好的细胞活性和更高的相关基因表达。结论： 直径为3.5cm的培养皿培养的细胞团相对更适合用来培养三维牙周膜细胞团,应用于牙周组织再生。 |
| 英文摘要: |
| Objective: The aim of the study was to assess the effect upon various aspects of periodontal ligament cell pellet culture influenced by diamter of the culture dishes used. Methods: Human periodontal ligament(PDL) cells were isolated,cultured and examined. Cell proliferation and adbesion were examined. PDL cells were cultured in different diamters(3.5cm,6cm,10cm) of the culture dishes respectively in DMEM medium with ascorbic acid. After 2 weeks,the cells in the dishes could form different sheets which could detached from the culture plate,then the sheets was cultured in a tube to produce a compact cell pellets. After the cell pellets were mature,a variety of techniques were applied to assess the viability of periodontal ligament cell pellets,including histological analysis,fluoresent assessment,MTT assay and polymerase chain reaction(PCR） to detect two target gene expression in the cultured PDL cells. Results: The viability and some other special features of PDL cell pellets in culture dishes of different sizes are distinct. PDL cells from culture dishes with small diameters showed high viability and a elevated expression level of two target gene. Conclusions: The results indicated PDL cell pellets cultured in 3.5cm diameter dishes possessed the highest viability compared with that cultured in 6 cm or 10 cm diameter dishes.Accordingly,a proper diameter for three-dimensional periodontal ligament cell pellet probably should be applied to the periodontal tissue reconstruction. |
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