The effect of human platelet lysate on the differentiation ability of human adipose-derived stem cells cultured on ECM-coated surfaces

Gao, Yan; Ku, Nien Ju; Sung, Tzu Cheng; Higuchi, Akon; Hung, Chi Sheng; Lee, Henry Hsin Chung; Ling, Qing; Cheng, Nai-Ming; Umezawa, Akihiro; Barro, Lassina; Burnouf, Thierry; Ye, Qingsong; Chen, Hao

doi:10.1039/c9tb01764j

Cited by 15 publications

(21 citation statements)

References 70 publications

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“…Human adipose-derived stem/stromal cells (ASCs) are a mesodermally derived multipotent stem cell type with the ability to differentiate into mesenchymal derived cell types, i.e. adipocytes, chondrocytes and osteocytes, and can also differentiate into cells from other germinal layers[ 1 , 2 ]. ASCs are currently used in various clinical trials [clinicaltrials.gov] owing to their regenerative ability, immune and inflammatory process modulation, and other properties such as neovascularization and apoptosis reduction [ 3 , 4 ].…”

Section: Introductionmentioning

confidence: 99%

Application of a deep learning-based image analysis and live-cell imaging system for quantifying adipogenic differentiation kinetics of adipose-derived stem/stromal cells

et al. 2021

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Section: Introductionmentioning

confidence: 99%

Application of a deep learning-based image analysis and live-cell imaging system for quantifying adipogenic differentiation kinetics of adipose-derived stem/stromal cells

et al. 2021

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“…The isolation process produces the stromal vascular fraction (SVF), a heterogenous cell population containing different cell types, including ASCs, endothelial precursor cells (EPCs), endothelial cells (ECs), lymphocytes, macrophages, smooth muscle cells, pericytes, and pre-adipocytes [Bora and Majumdar, 2017]. Culturing the SVF on a coatingfree plastic surface leads to ASC adherence and proliferation, a key characteristic of ASCs/MSCs, while culture on specific surfaces can lead to unwanted effects on e.g., the proliferation and initiation of differentiation of ASCs [Van Dijk et al, 2008;Somaiah et al, 2015;Bostancioglu et al, 2017;Gao et al, 2019]. Positive staining for CD73, CD90, and CD105 as well as negative staining for CD31 and CD45 are required to phenotypically define ASCs [Bourin et al, 2014].…”

Section: Introductionmentioning

confidence: 99%

Adipose-Derived Stromal/Stem Cell Culture: Effects of Different Concentrations of Human Platelet Lysate in Media

Rivera‐Ballesteros

Brooks

Haastrup

et al. 2020

Cells Tissues Organs

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Adipose-derived stromal/stem cells (ASCs) are being tested as a possible treatment for a wide range of diseases to exploit the immunomodulatory and regenerative potential demonstrated in vitro. Pooled human platelet lysate (pHPL) has replaced fetal bovine serum (FBS) as the preferred growth supplement because of its xeno-free origin and improved cell proliferation. Much has been done toward reducing the concentration of pHPL required when expanding ASCs. However, little is known on how increasing the concentration of pHPL affects ASC potency, which could lead to changes with possible beneficial applications. This study investigated the effect of 5, 10, or 20% pHPL in culture media on ASC proliferation and phenotypic marker expression, including chemokine receptors CXCR2, CXCR3, CXCR4, and VLA-4. Adipogenic and osteogenic properties, as well as immunosuppressive properties, including the ability to induce indoleamine-pyrrole 2,3-dioxygenase 1 (IDO1) and suppress T cell proliferation, were also examined. We observed a significant increase in cell yield (approximately 2-fold) and a corresponding reduction in population doubling time and cell volume when doubling the concentration of pHPL in the growth media. ASCs maintained expression of phenotypic surface markers CD73, CD90, and CD105 and were negative for CD45 and CD31. The ability to induce IDO1 and suppress T cell proliferation was observed as well. Adipogenesis and osteogenesis, however, seem to be increased at higher concentrations of pHPL (20% > 10% > 5%), while expression of chemokine receptors CXCR2 and CXCR3 was lower. In conclusion, increasing the pHPL concentration to 20% could be used to optimize culture conditions when producing cells for clinical treatments and may even be used to enhance beneficial ASC properties depending on the desired therapeutic effect.

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“…The expression of cell surface markers and adipogenic/osteogenic differentiation capabilities were maintained after replacing FBS with HPL for ASC culture. In a collaborative work, we have previously shown that HPL promoted human ASC differentiation into osteoblasts and chondrocytes compared to FBS (Gao et al, 2019). With 5% HPL employed in this study, the proliferative activity of ASCs was significantly increased relative to 10% FBS.…”

Section: Discussionmentioning

confidence: 50%

“…The highly stimulatory effect of HPL on cell proliferation enabled large-scale expansion of ASC for clinical use, yielding a great advantage when compared with the standard FBS culture method (Cowper et al, 2019;Gao et al, 2019). Besides enhancing the proliferative activity, HPL was also shown to promote ASC differentiation into osteoblasts and chondrocytes compared to FBS (Gao et al, 2019). HPL was suggested to alter the ASC behavior and function via multiple signaling pathways (Kakudo et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Influence of Human Platelet Lysate on Extracellular Matrix Deposition and Cellular Characteristics in Adipose-Derived Stem Cell Sheets

Cheng

Lee

et al. 2020

Front. Cell Dev. Biol.

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Adipose-derived stem cell (ASC) is a valuable source of cell therapy. By stimulating extracellular matrix (ECM) secretion, ASC sheets can be fabricated with enhanced regenerative capabilities. In recent years, human platelet lysate (HPL) provides an attractive alternative to fetal bovine serum (FBS) for the ex vivo expansion of ASCs for clinical use. However, the effect of HPL on ASC sheet formation has not been previously determined. In this study, we compared ECM composition and cellular characteristics of ASC sheets cultured in growth medium supplemented with either FBS or HPL. HPL supplement significantly enhanced ASC proliferation without obvious change in the expression pattern of cell surface markers. We found that culturing ASCs with HPL rendered thicker cell sheets with significantly more ECM deposition, including collagen and fibronectin. Proteomic analysis of the FBS or HPL-cultured cell sheets showed diversity in ECM composition. HPL-cultured ASC sheets exhibited up-regulation of interleukin-6 and an anti-inflammatory cytokine, C1q/tumor necrosis factor-related protein-3. Conditioned medium of HPL-cultured ASC sheets significantly enhanced fibroblast migration and tube formation of endothelial cells in vitro, while it inhibited the migration of macrophages toward stimulated macrophages in vitro. TGF-β1-stimulated fibroblasts cultured in ASC sheet-conditioned medium showed down-regulation of α-SMA and TGF-β1. By adding an anti-hepatocyte growth factor (HGF) neutralizing antibody in conditioned medium, we indicated that an anti-fibrosis effect of HPL-cultured ASC sheets is partially mediated through the increased secretion of HGF. Moreover, chick embryo chorioallantoic membrane (CAM) assay showed comparable capillary density after applying either FBS or HPL-cultured ASC sheets, both of which were significantly higher than the control. In conclusion, robust ECM formation with altered ECM composition was noted in ASC sheets cultured in HPL-supplemented medium. Their immunomodulatory and pro-angiogenesis capabilities were largely maintained. Our findings paved the way to elucidate the potential of HPL-cultured ASC sheets for clinical application in tissue regeneration.

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The effect of human platelet lysate on the differentiation ability of human adipose-derived stem cells cultured on ECM-coated surfaces

Abstract: Synchronized effects of cell culture materials and cell culture medium on osteoblast (left) and chondrocyte (right) differentiation were observed.

Cited by 15 publications

References 70 publications

Application of a deep learning-based image analysis and live-cell imaging system for quantifying adipogenic differentiation kinetics of adipose-derived stem/stromal cells

Application of a deep learning-based image analysis and live-cell imaging system for quantifying adipogenic differentiation kinetics of adipose-derived stem/stromal cells

Adipose-Derived Stromal/Stem Cell Culture: Effects of Different Concentrations of Human Platelet Lysate in Media

Influence of Human Platelet Lysate on Extracellular Matrix Deposition and Cellular Characteristics in Adipose-Derived Stem Cell Sheets

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