CAJ | 학술논문

　　背景：冻存是保证干细胞移植治疗的关键步骤之一。传统的冻存是将细胞直接置于冻存液中进行保存，然而冻存液中二甲基亚砜虽能减少细胞复苏过程中冰晶对细胞膜产生的机械性损伤，但同时又对细胞具有毒性作用，直接影响细胞生存状态，不利于临床移植治疗。目的：寻找适宜牙周膜干细胞体外扩增的牙周周组织冻存的最佳方案。方法：收集健康人牙，刮取牙周组织后将其平均分为3等份，随机分为新鲜组、5%二甲基亚砜组和10%二甲基亚砜组，后2组分别以体积分数5%和10%二甲基亚砜添加冻存1个月后提取牙周膜干细胞。新鲜组直接提取牙周膜干细胞。结果与结论：5%二甲基亚砜组原代细胞游出组织团块所需时间和细胞收获量虽不及新鲜培养组，但却明显优于10%二甲基亚砜组(P<0.05)。新鲜组、5%二甲基亚砜组和10%二甲基亚砜组第1代牙周膜干细胞克隆形成率、活细胞比率、第3代牙周膜干细胞BrdU细胞增殖能力、MTT细胞生长曲线和牙周膜干细胞表面标志物表达差异没有显著性意义(P>0.05)。提示5%二甲基亚砜添加冻存体系不仅能比10%二甲基亚砜添加的普通冻存体系明显缩短牙周膜干细胞体外扩增所需时间，增加细胞收获量同时还能保持细胞基本生物学特性，降低二甲基亚砜的总体用量和其在反复冻存复苏细胞过程中对细胞造成的直接损伤，为未来更加安全的实施临床移植治疗提供了保障，是供体组织储存新的选择。
　　배경：동존시보증간세포이식치료적관건보취지일。전통적동존시장세포직접치우동존액중진행보존，연이동존액중이갑기아풍수능감소세포복소과정중빙정대세포막산생적궤계성손상，단동시우대세포구유독성작용，직접영향세포생존상태，불리우림상이식치료。목적：심조괄의아주막간세포체외확증적아주주조직동존적최가방안。방법：수집건강인아，괄취아주조직후장기평균분위3등빈，수궤분위신선조、5%이갑기아풍조화10%이갑기아풍조，후2조분별이체적분수5%화10%이갑기아풍첨가동존1개월후제취아주막간세포。신선조직접제취아주막간세포。결과여결론：5%이갑기아풍조원대세포유출조직단괴소수시간화세포수획량수불급신선배양조，단각명현우우10%이갑기아풍조(P<0.05)。신선조、5%이갑기아풍조화10%이갑기아풍조제1대아주막간세포극륭형성솔、활세포비솔、제3대아주막간세포BrdU세포증식능력、MTT세포생장곡선화아주막간세포표면표지물표체차이몰유현저성의의(P>0.05)。제시5%이갑기아풍첨가동존체계불부능비10%이갑기아풍첨가적보통동존체계명현축단아주막간세포체외확증소수시간，증가세포수획량동시환능보지세포기본생물학특성，강저이갑기아풍적총체용량화기재반복동존복소세포과정중대세포조성적직접손상，위미래경가안전적실시림상이식치료제공료보장，시공체조직저존신적선택。
cryopreservation periodontal tissue. METHODS:Periodontal tissue was scraped from healthy human teeth and divided them into three equal parts:fresh group, harvesting periodontal ligament stem cel s from fresh tissue;5%dimethyl sulfoxide group, 5%dimethyl sulfoxide added into the cryopreservation system;10%dimethyl sulfoxide group, 10%dimethyl sulfoxide added into the cryopreservation system. One month later, periodontal ligament stem cel s were extracted from the cryopreserved periodontal ligament from the latter two groups. RESULTS AND CONCLUSION:In the time that cel s swam out of tissue mass and cel harvest amount, the 5%dimethyl sulfoxide group was inferior to the fresh group but better than the 10%dimethyl sulfoxide group (P<0.05). No differences were found among the three groups in the fol owing aspects (P>0.05):colony formation rate of passage 1 periodontal ligament stem cel s, cel survival rate, proliferation ability of passage 3 periodontal ligament stem cel s, cel growth curve and surface marker expression of periodontal ligament stem cel s. The results suggest that the 5%dimethyl sulfoxide added cryopreserved system for periodontal ligament tissue cannot only shorten the time of periodontal ligament stem cel s amplification in vitro, ensure cel harvest and maintain basic cel ular biological characteristics, but also reduce the total amount of the dimethyl sulfoxide and the direct damage to the cel s caused by repeatedly frozen cel s, thereby providing a more secure guarantee for the future implementation of clinical transplantation therapy. So, the 5%dimethyl sulfoxide added cryopreserved system may be the new selection for donor tissue storage.