Zare Sona, Ahmadi Rahim, Mohammadnia Abdolreza, Nilforoushzadeh Mohammad Ali, Mahmoodi Minoo
Department of Biology, Hamedan Branch, Islamic Azad University, Hamedan, Iran.
Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Lasers Med Sci. 2021 May 1;12:e18. doi: 10.34172/jlms.2021.18. eCollection 2021.
The efficiency of stem cell isolation, culture, and biological characterization techniques for treatment is facing serious challenges. The purpose of this study was to provide a protocol for isolation and culture of three types of mesenchymal stem cells (MSCs) derived from the human placenta, amniotic membrane, and umbilical cord with high efficiency used for cell therapy. During this experimental laboratory study, 10 complete placenta samples were prepared from cesarean section mothers. The protocol for isolation and culture of mesenchymal cells from the placenta tissue, umbilical cord, and amniotic membrane was enzymatically optimized. The morphological features of mesenchymal cells were investigated using an inverted microscope and their biological features were measured using flow cytometry. The differentiation potential of the cells was evaluated by measuring their differentiation capacity into osteocytes and adipocytes. The absorption and reflectance features of the cells were recorded by optical spectroscopy. Finally, the data were statistically analyzed. The expression of CD44, CD73, CD90 and CD29 markers in human placenta tissue-derived cells was significant. CD14, CD34 and CD45 markers were not expressed or were slightly expressed. These cells were highly viable and successfully differentiated into osteocytes and adipocytes. MSCs absorbed more light than visible light by showing light absorption peaks at wavelengths of about 435 and 550 nm. The protocol used in this study for isolation and culture of human placenta tissue-derived MSCs had significant efficiency for the production of MSCs for use in cell therapy and tissue engineering.
用于治疗的干细胞分离、培养及生物学特性鉴定技术的效率正面临严峻挑战。本研究的目的是提供一种高效分离和培养源自人胎盘、羊膜和脐带的三种间充质干细胞(MSC)的方案,用于细胞治疗。在这项实验性实验室研究中,从剖宫产母亲处获取了10份完整的胎盘样本。对从胎盘组织、脐带和羊膜中分离和培养间充质细胞的方案进行了酶法优化。使用倒置显微镜研究间充质细胞的形态特征,并使用流式细胞术测量其生物学特征。通过测量细胞向骨细胞和脂肪细胞的分化能力来评估细胞的分化潜能。通过光谱法记录细胞的吸收和反射特征。最后,对数据进行统计分析。人胎盘组织来源细胞中CD44、CD73、CD90和CD29标志物的表达显著。CD14、CD34和CD45标志物未表达或表达微弱。这些细胞具有高活力,并成功分化为骨细胞和脂肪细胞。MSC在约435和550nm波长处显示光吸收峰,对可见光的吸收多于可见光。本研究中用于分离和培养人胎盘组织来源MSC的方案在生产用于细胞治疗和组织工程的MSC方面具有显著效率。