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背景:研究发现富血小板血浆在促进骨再生、软组织修复以及组织工程研究中种子细胞的增殖分化方面有着重要的作用。目的:观察不同制备方法、不同体积分数富血小板血浆对体外培养大鼠毛囊干细胞增殖的影响。方法:联合采用显微分离技术,两步酶法及差速贴壁法获取纯度较高、生长状态较好的第3代大鼠毛囊干细胞。采用2次离心法和3次离心法获取大鼠富血小板血浆,分别按1.0%,2.0%,4.0%不同浓度稀释于K-SFM培养基中,以加入含不同体积分数富血小板血浆培养基组为实验组,并以未加入富血小板血浆的培养基为对照。结果与结论:联合采用显微分离技术,两步酶法及差速贴壁法能获取纯度较高的毛囊干细胞。培养前2d各实验组与对照组的比较差异无显著性意义(P>0.05),实验组加入富血小板血浆干预2d后,毛囊干细胞出现较高的增殖活性,第4天开始进入生长对数期,细胞第5天即达到70%-80%汇合,培养六七天细胞进入平台期。高剂量富血小板血浆组对细胞增殖效果优于低剂量组,体积分数相同的情况下3次离心法制备所得富血小板血浆对毛囊干细胞增殖作用明显高于2次离心法,以加入3次离心法制备且体积分数为4.0%的富血小板血浆组毛囊干细胞增殖效果最明显。
BACKGROUND: Platelet-rich plasma has been found to play an important role in promoting the proliferation and differentiation of seed cells in bone regeneration, soft tissue repair, and tissue engineering. Objective: To observe the effects of different preparation methods and different volume fractions of platelet-rich plasma on the proliferation of rat hair follicle stem cells cultured in vitro. Methods: The third generation of rat hair follicle stem cells with higher purity and better growth status were obtained by microscopic separation, two - step enzymatic method and differential adhesion method. Rat platelet-rich plasma was obtained by 2-fold centrifugation and 3-fold centrifugation. The rat platelet-rich plasma was diluted to 1.0%, 2.0% and 4.0% respectively in K-SFM medium to add different concentrations of platelet- For the experimental group, and not added to platelet-rich medium as a control. RESULTS AND CONCLUSION: The combined use of microscopic separation technology, two-step enzymatic method and differential adhesion method can obtain high purity hair follicle stem cells. There was no significant difference between the experimental group and the control group 2 d before the culture (P> 0.05). The hair follicle stem cells showed higher proliferative activity after 2 days intervention with platelet-rich plasma in experiment group, , The cells reach the 70% -80% confluence on the fifth day, and the cells on the sixth and seventh days are brought into the plateau. High-dose platelet-rich plasma group had better effect on cell proliferation than low-dose group. When the volume fraction was the same, the platelet-rich plasma obtained from 3 times centrifugation had significantly higher proliferation of hair follicle stem cells than that of 2 times centrifugation, The proliferation of hair follicle stem cells with platelet-rich plasma was most obvious when the volume fraction was 4.0%.