联合培养法进行人牙周膜细胞的比较培养与鉴定

目的 比较消化法与组织块法联合培养与单纯组织块法两种方法获得人牙周膜细胞的成功率及细胞来源鉴定情况.方法 刮取因正畸需要而拔牙的牙周膜,分别进行组织块法和消化法与组织块法联合培养进行人牙周膜细胞的原代培养,并进行波形丝蛋白、角蛋白和碱性磷酸酶(ALP)的检测,以鉴定细胞来源.结果 组织块法在1～2周内有原代细胞游出.在第2或第3天,联合培养法消化游离出来的细胞开始贴壁,消化后的疏松状牙周膜亦可贴壁,且有细胞爬出.组织块法的成功率为27％,联合培养法成功率为70％.ABC免异组化法及ALP鉴定其为非牙龈来源的中胚层细胞.结论 联合培养法可显著提高原代细胞培养的成功率,并在短期内获得大量和相对纯化的实验用细胞.

Culture and Identification of Human Periodontal Ligment Cells by Combined Method

Objective To evaluate the success rate of obtaining human periodontal ligment cells(HPLCs) and cells identification in vitro.Methods We scraped the periodontal ligament from human teeth which were extracted from who need to orthodontic repair,and obtained the HPLCs by tissue block method and combined method of enzymolytic and tissue block.Cells were indentified through detecting vimentin,keratin and Alkaline Phosphatase(ALP).Results The cells emigrated from tissue block after 1-2 weeks in tissue block method,however cells attachment occurred after 2-3 days in combined method.The success rate of tissue block method and combined method was 27% and 70%,respectively.The cells cultured were indentified as non-gingival derived mesoblastemas by ABC immunohistochemistry and ALP method.Conclusion Combined culture method could improve success rate of primary culture cells apparently and obtain plenty of cells in short time.