Hassan Ghmkin, Kasem Issam, Soukkarieh Chadi, Aljamali Majd
Department of Microbiology and Biochemistry, Faculty of Pharmacy, Damascus University, Damascus, Syria.
Department of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria.
Int J Stem Cells. 2017 Nov 30;10(2):184-192. doi: 10.15283/ijsc17028.
Mesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated from umbilical cords and are therapeutically used because of their ability to differentiate into various types of cells, in addition to their immunosuppressive and anti-inflammatory properties. Fetal bovine serum (FBS), considered as the standard additive when isolating and culturing MSCs, has a major limitation related to its animal origin. Here, we employed a simple and economically efficient protocol to isolate MSCs from human umbilical cord tissues without using digestive enzymes and replacing FBS with umbilical cord blood serum (CBS).
MSCs were isolated by culturing umbilical cord pieces in CBS or FBS supplemented media. Expansion and proliferation kinetics of cells isolated by explant method in the presence of either FBS or CBS were measured, with morphology and multi-differentiation potential of expanded cells characterized by flow cytometry, RT-PCR, and immunofluorescence. MSCs maintained morphology, immunophenotyping, multi-differentiation potential, and self-renewal ability, with better proliferation rates for cells cultured in CBS compared to FBS supplement media.
We here present a simple, reliable and efficient method to isolate MSCs from umbilical cord tissues, where cells maintained proliferation, differentiation potential and immunophenotyping properties and could be efficiently expanded for clinical applications.
间充质干细胞（MSCs）是多能干细胞，可从脐带中分离得到，因其能够分化为多种类型的细胞，以及具有免疫抑制和抗炎特性而被用于治疗。胎牛血清（FBS）在分离和培养MSCs时被视为标准添加剂，但其动物来源存在重大局限性。在此，我们采用了一种简单且经济高效的方案，无需使用消化酶，用脐带血血清（CBS）替代FBS，从人脐带组织中分离MSCs。
将脐带片段在添加CBS或FBS的培养基中培养以分离MSCs。测量了在FBS或CBS存在下通过组织块法分离的细胞的扩增和增殖动力学，通过流式细胞术、RT-PCR和免疫荧光对扩增细胞的形态和多分化潜能进行了表征。MSCs保持了形态、免疫表型、多分化潜能和自我更新能力，与添加FBS的培养基相比，在CBS中培养的细胞增殖率更高。
我们在此提出了一种从脐带组织中分离MSCs的简单、可靠且高效的方法，所分离的细胞保持了增殖、分化潜能和免疫表型特性，并且能够有效地扩增以用于临床应用。
