Abstract
Human adipose-derived stromal cells (ADSCs) are receiving unprecedented attention as a potential cellular source for regenerative medicine-based therapies against various diseases and conditions. However, there still have significant issues concerning the translational development of ADSC-based therapies, such as its heterogeneity and being prone to aging. We developed a new simple and economical 3D semi-suspended expansion method in which 3D spheroids reside on an ADSC-derived self-feeder cell layer, producing cells with increased population homogeneity and strong stemness and ensuring that the proliferation and differentiation potency of the cells does not become notably reduced after at least ten passages in culture. To check the potential application of the 3D ADSC spheroids, we discovered that the combination of siEID3, which is a small interfering RNA of EP300 inhibitor of differentiation 3 (EID3), and laminin/poly-d-lysine matrix can rapidly result in trans-differentiation of the 3D spheroid cells to neural progenitor-like cells (NPLCs) in approximately 9 days in vitro. This approach provides a multidisciplinary tool for stem cell research and production in mesenchymal stem cell-related fields.
Highlights
Human adipose-derived stromal/stem cells (ADSCs) are isolated from the “stromalvascular fraction” (SVF) of subcutaneous adipose tissue [1]
We cultured different batches of ADSC according to this method, these ADSCs are formed three-dimensional spheres on the ADSC-derived self-feeder cell layer (Supplementary Figure S1)
Flow cytometry (FCM) showed that expression of surface markers, such as CD29, CD73, CD90, and CD105, recommended by the ISCT was greater than 98%, confirming the mesenchymal stem cell characteristics of the cells (Figure 2)
Summary
Human adipose-derived stromal/stem cells (ADSCs) are isolated from the “stromalvascular fraction” (SVF) of subcutaneous adipose tissue [1] Due to these cells’ ready availability, large number, and abundant sources in the body, the number of studies on and applications of ADSCs have increased dramatically in recent years, and ADSCs had been extensively used in regenerative medicine mainly due to their paracrine effect, immunomodulatory functions and, differentiation capacity [2]. ADSCs exhibit high heterogeneity, which probably mainly results from various factors including donor age, different organ, isolation procedures, or culture methods [4], which affect the features of ADSCs, such as proliferative capacity, differentiation potential, aging process, immunophenotype, and secretory ability [4,5] For this reason, the application of ADSCs for clinical treatment has been greatly restricted [6], and more efficient solutions are needed. It has been reported that the 3D culture of human ADSCs promotes cell yields, maintains stemness, and represents a promising strategy for cell expansion on industrial levels, with great potential for cell therapy and biotechnology [11,13,14]
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