Dehkordi Mehdi Banitalebi, Madjd Zahra, Chaleshtori Morteza Hashemzadeh, Meshkani Reza, Nikfarjam Laleh, Kajbafzadeh Abdol-Mohammad
Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran (IRI).
Oncopathology Research Center and Dep Pathology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran (IRI).
Cell Transplant. 2016 Jul;25(7):1287-1297. doi: 10.3727/096368915X582769.
Several reports have been published on the isolation, culture, and identification of mesenchymal stem cells (MSCs) from different anatomical regions of the umbilical cord (UC). UC is suitable for standardizing methods of MSC isolation because it is a uniform source with high MSC numbers. Although the UC is considered a medical waste after childbirth, ethical issues for its use must be considered. An increased demand for MSCs in regenerative medicine has made scientists prioritize the development of MSC isolation methods. Several research groups are attempting to provide a large number of high-quality MSCs. In this study, we present a modulated explant/enzyme method (MEEM) to isolate the maximum number of MSCs from the entire UC. This method was established for the isolation of MSCs from different anatomical regions of the UC altogether. We could retrieve 6 to 10 million MSCs during 8 to 10 days of primary culture. After three passages, we could obtain 8-10 × 10 cells in 28-30 days. MSCs isolated by this method express CD73, CD90, CD105, and CD44, but they do not express hematopoietic markers CD34 and CD45 or the endothelial marker CD31. The genes SOX2, OCT4, and NANOG are expressed in isolated MSCs. The capacity of these MSCs to differentiate into adipocytes and osteocytes highlights their application in regenerative medicine. This method is simple, reproducible, and cost efficient. Moreover, this method is suitable for the production of a large number of high-quality MSCs from an UC in less than a month, to be used for cell therapy in an 80-kg person.
关于从脐带（UC）不同解剖区域分离、培养和鉴定间充质干细胞（MSCs）的研究已有多篇报道。UC适合标准化MSCs分离方法，因为它是一个具有大量MSCs的统一来源。尽管UC在分娩后被视为医疗废物，但使用它时必须考虑伦理问题。再生医学对MSCs的需求增加，促使科学家优先开发MSCs分离方法。几个研究小组正在尝试提供大量高质量的MSCs。在本研究中，我们提出了一种改良的外植体/酶法（MEEM），以从整个UC中分离出最大数量的MSCs。该方法是为从UC的不同解剖区域共同分离MSCs而建立的。在原代培养的8至10天内，我们可以获得600万至1000万个MSCs。传代三次后，我们可以在28至30天内获得8 - 10×10个细胞。通过这种方法分离的MSCs表达CD73、CD90、CD105和CD44，但不表达造血标志物CD34和CD45或内皮标志物CD31。基因SOX2、OCT4和NANOG在分离的MSCs中表达。这些MSCs分化为脂肪细胞和骨细胞的能力突出了它们在再生医学中的应用。该方法简单、可重复且成本效益高。此外，这种方法适合在不到一个月的时间内从UC中生产大量高质量的MSCs，用于80公斤体重的人的细胞治疗。
