The Role of Glucose, Serum, and Three-Dimensional Cell Culture on the Metabolism of Bone Marrow-Derived Mesenchymal Stem Cells

Deorosan, Byron Anthony; Nauman, Eric A.

doi:10.4061/2011/429187

Cited by 37 publications

(22 citation statements)

References 59 publications

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“…Moreover, NANOG mRNA levels were intensely down-regulated in bMSC treated with the same concentration. Supplementation of FBS to culture media has been shown to increase proliferation of MSC [38]. The effect of FBS alters the metabolic state of MSC and may also affect the terminal differentiation of MSC.…”

Section: Discussionmentioning

confidence: 99%

Isolation and multilineage differentiation of bone marrow mesenchymal stem cells from abattoir-derived bovine fetuses

et al. 2013

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BackgroundMesenchymal stem cells (MSC) are multipotent progenitor cells localized in the stromal compartment of the bone marrow (BM). The potential of MSC for mesenchymal differentiation has been well documented in different animal models predominantly on rodents. However, information regarding bovine MSC (bMSC) is limited, and the differentiation potential of bMSC derived from fetal BM remains unknown. In the present study we sought to isolate bMSC from abattoir-derived fetal BM and to characterize the multipotent and differentiation potential under osteogenic, chondrogenic and adipogenic conditions by quantitative and qualitative analyses.ResultsPlastic-adherent bMSC isolated from fetal BM maintained a fibroblast-like morphology under monolayer culture conditions. These cells expressed high levels of MSC surface markers (CD73, CD90, and CD105) and low levels of hematopoietic surface markers (CD34 and CD45). Culture of bMSC under osteogenic conditions during a 27-day period induced up-regulation of the osteocalcin (OC) gene expression and alkaline phosphatase (ALPL) activity, and promoted mineralization of the matrix. Increasing supplementation levels of ascorbic acid to culture media enhanced osteogenic differentiation of bMSC; whereas, reduction of FBS supplementation compromised osteogenesis. bMSC increased expression of cartilage-specific genes aggrecan (ACAN), collagen 2A1 (COL2A1) and SRY (sex-determining region Y) box 9 (SOX9) at Day 21 of chondrogenic differentiation. Treatment of bMSC with adipogenic factors increased levels of fatty acid-binding protein 2 (AP2) mRNA and accumulation of lipid vacuoles after 18 days of culture. NANOG mRNA levels in differentiating bMSC were not affected during adipogenic culture; however, osteogenic and chondrogenic conditions induced higher and lower levels, respectively.ConclusionsOur analyses revealed the potential multilineage differentiation of bMSC isolated from abattoir-derived fetal BM. NANOG mRNA pattern in differentiating bMSC varied according to differentiation culture conditions. The osteogenic differentiation of bMSC was affected by ascorbic acid and FBS concentrations in culture media. The simplicity of isolation and the differentiation potential suggest that bMSC from abattoir-derived fetal BM are appropriate candidate for investigating MSC biology and for eventual applications for regenerative therapy.

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Section: Discussionmentioning

confidence: 99%

Isolation and multilineage differentiation of bone marrow mesenchymal stem cells from abattoir-derived bovine fetuses

et al. 2013

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“… Note . Literature values for bovine articular chondrocytes (Suits, 2006), hMSCs (Higuera et al, ; Pattappa, Heywood, de Bruijn, & Lee, ; Schop et al, ), and murine mesenchymal stem cells (Deorosan & Nauman, ) in static culture are included for comparison. Note that literature values for hAC/hMSC co‐culture in static conditions were not available, and literature values for bioreactor culture of hACs or hMSCs were also not available.…”

Section: Resultsmentioning

confidence: 99%

Osteoarthritic human chondrocytes proliferate in 3D co‐culture with mesenchymal stem cells in suspension bioreactors

Khurshid

Mulet‐Sierra

Adesida

et al. 2017

J Tissue Eng Regen Med

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Osteoarthritis (OA) is a painful disease, characterized by progressive surface erosion of articular cartilage. The use of human articular chondrocytes (hACs) sourced from OA patients has been proposed as a potential therapy for cartilage repair, but this approach is limited by the lack of scalable methods to produce clinically relevant quantities of cartilage-generating cells. Previous studies in static culture have shown that hACs co-cultured with human mesenchymal stem cells (hMSCs) as 3D pellets can upregulate proliferation and generate neocartilage with enhanced functional matrix formation relative to that produced from either cell type alone. However, because static culture flasks are not readily amenable to scale up, scalable suspension bioreactors were investigated to determine if they could support the co-culture of hMSCs and OA hACs under serum-free conditions to facilitate clinical translation of this approach. When hACs and hMSCs (1:3 ratio) were inoculated at 20,000 cells/ml into 125-ml suspension bioreactors and fed weekly, they spontaneously formed 3D aggregates and proliferated, resulting in a 4.75-fold increase over 16 days. Whereas the apparent growth rate was lower than that achieved during co-culture as a 2D monolayer in static culture flasks, bioreactor co-culture as 3D aggregates resulted in a significantly lower collagen I to II mRNA expression ratio and more than double the glycosaminoglycan/DNA content (5.8 vs. 2.5 μg/μg). The proliferation of hMSCs and hACs as 3D aggregates in serum-free suspension culture demonstrates that scalable bioreactors represent an accessible platform capable of supporting the generation of clinical quantities of cells for use in cell-based cartilage repair.

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“…Effects due to cell immobilization are often not considered, and many of the differences we observed between monolayer and 3D culture conditions may be due to the effects of cell–ECM interactions, such as cellular stress produced by the immobilization process (Schneider et al ., ; Sun et al ., ), changes in cellular metabolism (Constantinidis et al ., ; Simpson et al ., ) or mechanical stimulation (Constantinidis et al , ; Bilodeau and Mantovani, ). Because cellular metabolism is altered in 3D culture compared to monolayer culture (Figure ), this may have significant implications in the use of matrices, because metabolism affects all cell physiology (Deorosan and Nauman, ). Our results suggest that the different chemical compositions of the matrices do not a priori influence TGF β 3 secretion.…”

Section: Discussionmentioning

confidence: 99%

TGFβ3 secretion by three-dimensional cultures of human dental apical papilla mesenchymal stem cells

Somoza

Acevedo

Albornoz

et al. 2015

J Tissue Eng Regen Med

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Mesenchymal stem cells (MSCs) can be isolated from dental tissues, such as pulp and periodontal ligament; the dental apical papilla (DAP) is a less-studied MSC source. These dental-derived MSCs are of great interest because of their potential as an accessible source for cell-based therapies and tissue-engineering (TE) approaches. Much of the interest regarding MSCs relies on the trophic-mediated repair and regenerative effects observed when they are implanted. TGFβ3 is a key growth factor involved in tissue regeneration and scarless tissue repair. We hypothesized that human DAP-derived MSCs (hSCAPs) can produce and secrete TGFβ3 in response to micro-environmental cues. For this, we encapsulated hSCAPs in different types of matrix and evaluated TGFβ3 secretion. We found that dynamic changes of cell-matrix interactions and mechanical stress that cells sense during the transition from a monolayer culture (two-dimensional, 2D) towards a three-dimensional (3D) culture condition, rather than the different chemical composition of the scaffolds, may trigger the TGFβ3 secretion, while monolayer cultures showed almost 10-fold less secretion of TGFβ3. The study of these interactions is provided as a cornerstone in designing future strategies in TE and cell therapy that are more efficient and effective for repair/regeneration of damaged tissues. Copyright © 2015 John Wiley & Sons, Ltd.

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The Role of Glucose, Serum, and Three-Dimensional Cell Culture on the Metabolism of Bone Marrow-Derived Mesenchymal Stem Cells

Cited by 37 publications

References 59 publications

Isolation and multilineage differentiation of bone marrow mesenchymal stem cells from abattoir-derived bovine fetuses

Isolation and multilineage differentiation of bone marrow mesenchymal stem cells from abattoir-derived bovine fetuses

Osteoarthritic human chondrocytes proliferate in 3D co‐culture with mesenchymal stem cells in suspension bioreactors

TGFβ3 secretion by three-dimensional cultures of human dental apical papilla mesenchymal stem cells

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