In the present study, sample 2 maintained proliferative capacities even after 20 passages in SFM. stem cells (MSCs) possess the ability of self-renew and multipotency. These cells can replicate and have the potential to differentiate to bone, fat and cartilage tissues [1]. Dihydromyricetin (Ampeloptin) Numerous preclinical and clinical studies have exhibited that MSCs hold an alluring prospect as cellular therapies, based on their multipotency, hematopoietic-supporting and immunosuppressive abilities. MSC-based tissue-engineering approaches could treat patients with long bone defects [2, 3]. Co-transplantation of MSCs with HLA-disparate hematopoietic stem cells could accelerate lymphocyte recovery and reduce the risk of graft failure [4]. For the small effect on T-cell responses to pathogens, infusion of MSCs suppressed alloantigen-induced T-cell function and might be a promising therapy for graft-versus-host disease [5, 6]. MSCs, described as a very rare population in bone marrow by Friedenstein and colleagues [7], need to be expanded to achieve the amount required for administration. Several safety-related issues have been of wide concern in clinical applications of expanded MSCs. It is not clear whether MSCs could maintain genomic stability during expansion and whether injection of MSCs could lead to cancer culture in serum-containing culture [8]. Other studies on pluripotent stem cells have revealed that the number of chromosomes and the copy number of specific regions in the genome of embryonic stem cells or induced pluripotent stem cells could mutate in the process of expansion [9C14]. Similarly, copy number variation (CNV) was found Rabbit Polyclonal to GFP tag in adipose tissue-derived MSCs [15] after long-term culture, even though they did not undergo malignant transformation. Previous studies paid much attention to the safety issues of MSCs cultured in serum-containing medium (SCM) [16]. However, it is not desirable to prepare MSCs for clinical application in SCM. The utmost problem associated with bovine and human serum is the safety issue. Bovine serum might contain zoonotic viruses (including prion), which cannot be cleaned up during the process of preparing MSCs for clinical use. Human serum might contain undetectable pathogen, which could easily spread between human beings during stem cell transplantation. From this perspective, human serum is more dangerous than serum of animals. In recent years, some human serum or human platelet lysate products are solvent/detergent treated, which makes them much less likely to transmit an infectious disease, without deleting the risk completely [17]. In addition, Dihydromyricetin (Ampeloptin) serum is usually ill-defined, has a high degree of batch-to-batch variation, is usually hard to standardize and can harm the process control and stability of quality and production. Serum-free medium (SFM) is an ideal system for cellular therapy. MSCs expanded in SFM perform much better in quality control and stability. Many previous studies focused on increasing attachment and growth of MSCs in SFM [18]. Other studies evaluated the clinical application related biological characteristics of SFM-expanded MSCs [19, 20]. However, the safety and efficacy of MSCs cultured in SFM have not been well evaluated [21]. In this study we investigated whether human umbilical cord mesenchymal stem cells (hUC-MSCs) expanded in SFM change their biological characteristics and clinical safety-related issues, which included genome and transcriptome stability. Methods Growth characteristics of MSCs in serum-free medium hUC-MSCs derived from five different donors were isolated from Whartons jelly by enzymatic digestion [22] and frozen in a grasp cell bank after short-term expansion in SCM. This study is approved Dihydromyricetin (Ampeloptin) by the Institutional Review Board of the Chinese Academy of Medical Science and Peking Union Medical College. Umbilical cords were obtained following the ethical guidelines with written informed consent from donors. All experimental research of the scholarly research is at compliance using the Helsinki Declaration. After recovery through the get better at cell standard bank, hUC-MSCs had been cultured on the tissue culture surface area with SCM that included 10% fetal bovine serum (ExCell Bio, Shanghai, China) or on the chemically treated cell tradition surface area (CellBIND; Corning Integrated, Corning, NY, USA) having a chemically described SFM (MSCGM-CD; Lonza, Walkersville, MD, USA), at 37C and 5% skin tightening and. After achieving 90% confluence, hUC-MSCs had been subcultured and detached in a percentage of just one 1:3 until getting senescence. The proper time had a need to obtain confluence for each and every passage was recorded to calculate the population-doubling time. -galactosidase had been analyzed at past due passing by a mobile senescence assay package (Millipore, Billerica, MA, USA) following a manufacturers protocol. With this assay, senescent cells had been stained as a unique blue color. Differentiation of MSCs cultured in serum-free moderate For adipogenic and osteogenic differentiation, SFM-expanded.