摘要
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： Recent studies have found that the platelet-rich plasma has an important role in promoting bone regeneration, soft tissue repair and proliferation and differentiation of the seed cells in tissue engineering. OBJECTIVE： To observe the effect of different methods and different volume fractions of platelet-rich plasma on the proliferation of rat hair follicle stem cells. METHODS： Combination of microdissection and two-step enzyme digestion with differential adhesion methods were used to obtain the third generation of rat hair follicle stem cells with high purity and good growth state. The second centrifugation method and the third centrifugation method were used to obtain the rat platelet-rich plasma, and then diluted into the K-SFM medium with the volume fraction of 1.0%, 2.0% and 4.0%, while the platelet-rich plasma containing different volume fractions of medium was considered as the experimental group and the platelet-rich plasma without medium was used as a control. RESULTS AND CONCLUSION： Highly purified hair follicle stem cells could be obtained by microdissection, two-step enzyme digestion and differential adhesion methods. There was no significant difference between experimental group and control group at 2 days before culture （P 〉 0.05）; and then hair follicle stem cells in the experimental group showed higher proliferation activity after the platelet-rich plasma intervention for 2 days. After cultured for 4 days, the hair follicle stem cells grew into the growth logarithmic phase; and after 5 days, the cells reached to 70%-80% confluence, and then went into the platform after cultured for 6-7 days. The cell proliferation in the high dose platelet-rich plasma group was better than that in the low dose platelet-rich plasma group. Under the condition of the same volume fraction, the effect of the platelet-rich plasma obtained by third centrifugation method on the proliferation of hair follicle stem cells was significantly greater than that obtained by the second centrifugation metho
出处
《中国组织工程研究》
CAS
CSCD
2012年第45期8501-8505,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81160088)
新疆维吾尔自治区自然科学基金项目(2009211B22)~~
