Effect of autologous platelet-rich plasma on directional induced differentiation of rabbit bone marrow mesenchymal stem cells

中华医学美学美容杂志 Pub Date : 2019-04-15 DOI:10.3760/CMA.J.ISSN.1671-0290.2019.02.020

Ma Zhibing, Fengying Yuan, P. Jin, A. Zhang, Chang-bo Tao, Xue-yang Li, Caiqi Shen, Yanping Guo

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Abstract

Objective To study the cell morphology and differentiation efficiency when rabbit bone marrow mesenchymal stem cells (BMSCs) were induced osteogenic differentiation as culturing by autologous platelet-rich plasma (PRP) instead of serum, and to explore a new method of inducing BMSCs osteogenic differentiation. Methods The PRP was prepared by arterial blood of rabbit. Punctured and The bone marrow was sampled from rabbit＇s iliac bone, and BMSCs were collected, which divided into PRP group, fetal calf serum (FBS) group and serum-free control group, and cultured in 10% autologous PRP, 10% FBS and serum-free respectively, combined with DMEM-F12 medium. The second generation cells were divided into experimental and control groups. The experimental groups＇ medium was added dexamethasone, β-sodium glycerophosphate and ascorbic acid, and the control groups went on. The cell morphological difference of each group was Observed between anterior and after inducing differentiation, and compared between each group. Results BMSCs of PRP and FBS groups grew quickly, presented like fusiform form before induction, and increasd in volume, became a triangle, polygonal and round form gradually after osteogenic induction. Cells of PRP and FBS groups aggregated spontaneously and multilayered, and formed calcium nodules and bone-like structure after induced 7 days averagely, which could be stained red by alizarin red S; cells of serum-free groups were induced 14 days averagely, only three samples showed osteogenesis performance. Cells of PRP and FBS groups differentiation efficiency was superior to serum-free groups when inducd 20 days, the difference was statistically significant (P 0.05). Conclusions Autologous PRP could be used to proliferate and induce osteogenic differentiation of BMSCs instead of serum. Key words: Rabbits; Bone marrow; Stromal cells; Stem cells; Platelet-rich plasma; Osteoblasts; Cell Differentiation

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自体富血小板血浆对兔骨髓间充质干细胞定向诱导分化的影响

目的研究自体富血小板血浆(PRP)代替血清诱导兔骨髓间充质干细胞(BMSCs)成骨分化时的细胞形态和分化效率，探索一种诱导BMSCs成骨分化的新方法。方法采用家兔动脉血制备PRP。取兔髂骨穿刺骨髓，采集骨髓间充质干细胞，分为PRP组、胎牛血清(FBS)组和无血清对照组，分别在10%自体PRP、10%胎牛血清和无血清中培养，并结合DMEM-F12培养基。第二代细胞分为实验组和对照组。实验组在培养液中加入地塞米松、β-甘油磷酸钠和抗坏血酸，对照组继续。观察各组细胞分化前和诱导分化后的形态差异，并比较各组间的差异。结果PRP组和FBS组骨髓间充质干细胞生长迅速，诱导前呈梭状，成骨诱导后体积逐渐增大，逐渐变成三角形、多边形和圆形。PRP组和FBS组细胞自发聚集，呈多层状，平均诱导7 d后形成钙结节和骨样结构，茜素红S染色;无血清组细胞平均诱导14 d，只有3个样本显示成骨能力。诱导20 d时，PRP组和FBS组细胞分化效率优于无血清组，差异有统计学意义(p0.05)。结论自体PRP可替代血清诱导骨髓间充质干细胞增殖和成骨分化。关键词:家兔;骨髓;基质细胞;干细胞;富含血小板血浆;成骨细胞;细胞分化

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来源期刊

中华医学美学美容杂志

自引率

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发文量

4641

期刊介绍： "Chinese Journal of Medical Aesthetics and Cosmetology" is a high-end academic journal focusing on the basic theoretical research and clinical application of medical aesthetics and cosmetology. In March 2002, it was included in the statistical source journals of Chinese scientific and technological papers of the Ministry of Science and Technology, and has been included in the full-text retrieval system of "China Journal Network", "Chinese Academic Journals (CD-ROM Edition)" and "China Academic Journals Comprehensive Evaluation Database". Publishes research and applications in cosmetic surgery, cosmetic dermatology, cosmetic dentistry, cosmetic internal medicine, physical cosmetology, drug cosmetology, traditional Chinese medicine cosmetology and beauty care. Columns include: clinical treatises, experimental research, medical aesthetics, experience summaries, case reports, technological innovations, reviews, lectures, etc.