Enhancing proliferation and optimizing the culture condition for human bone marrow stromal cells using hypoxia and fibroblast growth factor-2

Lee, Jung Seok; Kim, Seul Ki; Jung, Byung Joo; Choi, Seong Bok; Choi, Eun Young; Kim, Chang Sung

doi:10.1016/j.scr.2018.01.010

Cited by 23 publications

(23 citation statements)

References 27 publications

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“…It has been well-established that physioxic conditions enhance the anabolic response in MSC chondrogenesis [15,37,39,40,41,42,43,51,52]. Anderson et al (2016) showed that the reaction under physioxia is related to the GAG production under hyperoxia.…”

Section: Discussionmentioning

confidence: 99%

Physioxia Has a Beneficial Effect on Cartilage Matrix Production in Interleukin-1 Beta-Inhibited Mesenchymal Stem Cell Chondrogenesis

Pattappa

Schewior

Hofmeister

et al. 2019

Cells

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Osteoarthritis (OA) is a degenerative condition that involves the production of inflammatory cytokines (e.g., interleukin-1β (IL-1β), tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6)) that stimulate degradative enzymes, matrix metalloproteinases (MMPs) and aggrecanases (ADAMTS) resulting in articular cartilage breakdown. The presence of interleukin-1β (IL-1β) is one reason for poor clinical outcomes in current cell-based tissue engineering strategies for treating focal early osteoarthritic defects. Mesenchymal stem cells (MSCs) are a potential cell source for articular cartilage regeneration, although IL-1β has been shown to inhibit in vitro chondrogenesis. In vivo, articular chondrocytes reside under a low oxygen environment between 2–5% oxygen (physioxia) and have been shown to enhance in vitro MSC chondrogenic matrix content with reduced hypertrophic marker expression under these conditions. The present investigation sought to understand the effect of physioxia on IL-1β inhibited MSC chondrogenesis. MSCs expanded under physioxic (2% oxygen) and hyperoxic (20%) conditions, then chondrogenically differentiated as pellets in the presence of TGF-β1 and either 0.1 or 0.5 ng/mL IL-1β. Results showed that there were donor variations in response to physioxic culture based on intrinsic GAG content under hyperoxia. In physioxia responsive donors, MSC chondrogenesis significantly increased GAG and collagen II content, whilst hypertrophic markers were reduced compared with hyperoxia. In the presence of IL-1β, these donors showed a significant increase in cartilage matrix gene expression and GAG content relative to hyperoxic conditions. In contrast, a set of MSC donors were unresponsive to physioxia and showed no significant increase in matrix production independent of IL-1β presence. Thus, physioxia has a beneficial effect on MSC cartilage matrix production in responsive donors with or without IL-1β application. The mechanisms controlling the MSC chondrogenic response in both physioxia responsive and unresponsive donors are to be elucidated in future investigations.

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Section: Discussionmentioning

confidence: 99%

Physioxia Has a Beneficial Effect on Cartilage Matrix Production in Interleukin-1 Beta-Inhibited Mesenchymal Stem Cell Chondrogenesis

Pattappa

Schewior

Hofmeister

et al. 2019

Cells

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“…With this as a caveat, the majority of studies of MSC chondrogenesis either in pellet or scaffold cultures indicates an increase in chondrogenic genes ( COL2A1 and ACAN ) and matrix formation under physioxia [33,41,42,43,47,49,50,52,55,62,71,72,76,77,78,79,82,86,87,88,90,92,93,95,97]. Furthermore, it has also been shown that physioxia is a more potent promoter of chondrogenesis than dynamic (compressive) loading with no synergistic effect upon combining the parameters [78,98].…”

Section: Physioxia and Msc Chondrogenesismentioning

confidence: 99%

“…MSC that have been pre-expanded or preconditioned under physioxia prior to subsequent chondrogenesis under the same conditions demonstrated an enhancement in matrix deposition and a reduction in collagen X and other hypertrophy markers compared with non-preconditioned chondrogenic MSCs differentiated under physioxia (summarised in Table 2) [41,42,43,45,46,48,49,52,62,89,90,91,93,94,95,97]. Martin-Rendon et al (2007) showed that preconditioned MSCs had higher SOX9 gene expression and subsequent pellet wet weight [89].…”

Section: Physioxia and Msc Chondrogenesismentioning

confidence: 99%

The Importance of Physioxia in Mesenchymal Stem Cell Chondrogenesis and the Mechanisms Controlling Its Response

Pattappa

Johnstone

Zellner

et al. 2019

IJMS

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Articular cartilage covers the surface of synovial joints and enables joint movement. However, it is susceptible to progressive degeneration with age that can be accelerated by either previous joint injury or meniscectomy. This degenerative disease is known as osteoarthritis (OA) and it greatly affects the adult population. Cell-based tissue engineering provides a possible solution for treating OA at its earliest stages, particularly focal cartilage lesions. A candidate cell type for treating these focal defects are Mesenchymal Stem Cells (MSCs). However, present methods for differentiating these cells towards the chondrogenic lineage lead to hypertrophic chondrocytes and bone formation in vivo. Environmental stimuli that can stabilise the articular chondrocyte phenotype without compromising tissue formation have been extensively investigated. One factor that has generated intensive investigation in MSC chondrogenesis is low oxygen tension or physioxia (2–5% oxygen). In vivo articular cartilage resides at oxygen tensions between 1–4%, and in vitro results suggest that these conditions are beneficial for MSC expansion and chondrogenesis, particularly in suppressing the cartilage hypertrophy. This review will summarise the current literature regarding the effects of physioxia on MSC chondrogenesis with an emphasis on the pathways that control tissue formation and cartilage hypertrophy.

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“…Combined delivery of FGF-2, transforming growth factor-β1, and adipose-derived stem cells from an engineered periosteum to a critical-sized mouse femur defect yielded improved bone allograft healing (23). The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing human bone marrow-derived stem cells to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential (24). The effects of temporal manipulation were achieved by applying FGF-2 and transforming growth factor-β3 on the derivation of proliferative chondrocytes from mesenchymal stem cells (25).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

Tae

Park

2019

Exp Ther Med

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Fibroblast growth factor-2 (FGF-2) is reported to have various functions and is considered a key human mesenchymal stem cell mitogen, often supplemented to increase human mesenchymal stem cell growth rates. The purpose of this study was to evaluate the effects of FGF-2 on cellular viability and osteogenic differentiation using three-dimensional cell spheroids of stem cells. Three-dimensional cell spheroids were fabricated using concave silicon elastomer-based microwells in the presence of FGF-2 at concentrations of 0, 30, 60 and 90 ng/ml. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit-8 assay. Alkaline phosphatase activity and Alizarin Red S staining were used to assess osteogenic differentiation. Spheroids were well formed in silicon elastomer-based concave microwells on Day 1. The average spheroid diameters at Day 1 for FGF-2 at 0, 30, 60 and 90 ng/ml were 202.2±3.0, 206.6±22.6, 208.8±6.8 and 196.6±26.7 µm, respectively (P>0.05). The majority of the cells in the cell spheroids emitted green fluorescence. The relative Cell Counting Kit-8 assay values for FGF-2 at 0, 30, 60 and 90 ng/ml at Day 1 were 100.0±5.5, 101.8±8.8, 99.2±4.8 and 103.4±9.6% (P>0.05). The addition of FGF-2 at 60 ng/ml concentration produced the highest value for alkaline phosphatase activity. Mineralized extracellular deposits were evenly observed in each group, and the highest value was identified for FGF-2 groups at 60 ng/ml concentration for Alizarin Red S staining. Based on these findings, it was concluded that FGF-2 may increase alkaline phosphatase activity or Alizarin Red S staining, and further studies are needed to fully elucidate the mechanisms of FGF-2.

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Enhancing proliferation and optimizing the culture condition for human bone marrow stromal cells using hypoxia and fibroblast growth factor-2

Cited by 23 publications

References 27 publications

Physioxia Has a Beneficial Effect on Cartilage Matrix Production in Interleukin-1 Beta-Inhibited Mesenchymal Stem Cell Chondrogenesis

Physioxia Has a Beneficial Effect on Cartilage Matrix Production in Interleukin-1 Beta-Inhibited Mesenchymal Stem Cell Chondrogenesis

The Importance of Physioxia in Mesenchymal Stem Cell Chondrogenesis and the Mechanisms Controlling Its Response

Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

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