Derivation and Characterization of Mesenchymal Stem Cells from iPS Cells
Mesenchymal stem/stromal cells (MSCs) show excellent therapeutic potentials in many preclinical studies and clinical trials. However, the clinical application of conventional tissue-derived MSCs faces challenges of limited scalability and high donor varia
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Derivation and Characterization of Mesenchymal Stem Cells from iPS Cells Qingguo Zhao and Fei Liu Abstract Mesenchymal stem/stromal cells (MSCs) show excellent therapeutic potentials in many preclinical studies and clinical trials. However, the clinical application of conventional tissue-derived MSCs faces challenges of limited scalability and high donor variations. To address these challenges, we established a protocol for deriving and characterizing MSCs from human induced pluripotent stem cells (iPSCs) with a theoretically limitless expandability. The iPSC-MSCs show biological properties comparable to or better than early passage bone marrow MSCs and can be scaled up to huge amounts with uniform properties. Key words Mesenchymal stem/stromal cells, Induced pluripotent stem cells, Derivation, Characterization, Scalability
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Introduction Mesenchymal stem/stromal cells (MSCs) are multipotent adult stem cells that can differentiate into osteoblasts, chondrocytes, and adipocytes in vitro and in vivo [1]. A large amount of reports demonstrate therapeutic effects of MSCs in animal models and clinical trials of various degenerative diseases and inflammatory diseases [2]. Conventionally, MSCs are isolated from various tissues including bone marrow, adipose tissue, and umbilical cord [3]. However, the translational research and clinical application of tissue-derived MSC face many challenges: they have limited proliferation potentials and lose some important biological functions after prolonged expansion [4]; they are highly heterogeneous due to inconsistent isolation and culture procedures and donor differences in the age, genetic traits, and physiological conditions [5, 6]. Consequently, the standardization of tissue-derived MSCs is difficult, and the data generated in different laboratories are difficult to compare [7]. Therefore, more consistent and expandable MSCs are in great need. To overcome these limitations of tissue-derived MSCs, we derived MSCs from induced pluripotent stem cells (iPSCs) that have a theoretically unlimited expansion potential. Our iPSC-
Qingguo Zhao and Fei Liu
MSCs express classical surface markers of MSCs; are capable of differentiation into osteocytes, chondrocytes, and adipocytes; can home to various cancers with efficiencies comparable to earlypassage MSCs; and do not form teratoma in immune-compromised mice as iPSCs do [8]. Our derivation protocol is easily scalable, and we have established a bank of iPSC-MSCs with consistent biological properties [9, 10]. Moreover, our iPSC-MSCs and their extracellular vesicles are capable of repressing inflammation and modulating immune responses [11, 12], while our iPSC-MSCs and their matrix show powerful osteoregenerative capabilities [13]. Here we describe our detailed protocols for deriving and characterizing mesenchymal stem cells from human induced pluripotent stem cells.
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Materials Prepare all solutions using sterile ultrapure water (autoclaved deionized water with 18 MΩ-cm at 25 C) and/or sterile cell culture grade reagents in a biosafe
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