Ex vivo enrichment of mesenchymal cell progenitors by fibroblast growth factor 2

Bianchi, Giordano; Banfi, Andrea; Mastrogiacomo, Maddalena; Notaro, Rosario; Luzzatto, Lucio; Cancedda, Ranieri; Quarto, Rodolfo

doi:10.1016/s0014-4827(03)00138-1

Cited by 337 publications

(300 citation statements)

References 42 publications

Supporting

Mentioning

279

Contrasting

Unclassified

Order By: Relevance

“…In fact, removal of FGF-2 at different time points during serial replating always caused the disappearance of NAMP and its addition could not reinduce them at a later stage. This is in contrast to the effects on CFU-f, where FGF-2 induces, instead, an immediate and permanent selection of a subset of pluripotent mesenchymal precursors, even after a short exposure or when supplemented only after several days of culture [32]. The different biological response to FGF-2 reinforces the concept that NAMP are a distinct class of progenitors and not a proliferating fraction of the adherent CFU-f.…”

Section: Discussionmentioning

confidence: 73%

See 1 more Smart Citation

Fibroblast Growth Factor-2 Maintains a Niche-Dependent Population of Self-Renewing Highly Potent Non-adherent Mesenchymal Progenitors Through FGFR2c

et al. 2012

Self Cite

View full text Add to dashboard Cite

Bone marrow (BM) mesenchymal stem/stromal cells (MSC) are a heterogeneous population of multipotent progenitors currently under investigation for a variety of applications in regenerative medicine. While self-renewal of stem cells in different tissues has been demonstrated to be regulated by specialized microenvironments called niches, it is still unclear whether a self-renewing niche also exists for MSC. Here, we show that primary human BM cultures contain a population of intrinsically non-adherent mesenchymal progenitors (NAMP) with features of more primitive progenitors than the initially adhering colonyforming units-fibroblast (CFU-f). In fact, NAMP could generate an adherent progeny: (a) enriched with early mesenchymal populations (CD1461, SSEA-11, and SSEA-41); (b) with significantly greater proliferation and multilineage differentiation potential in vitro; and (c) capable of threefold greater bone formation in vivo than the corresponding CFU-f. Upon serial replating, NAMP were able to regenerate and expand in suspension as non-adherent clonogenic progenitors, while also giving rise to an adherent progeny. This took place at the cost of a gradual loss of proliferative potential, shown by a reduction in colony size, which could be completely prevented when NAMP were expanded on the initially adhering BM fraction. Mechanistically, we found that NAMP crucially depend on fibroblast growth factor (FGF)-2 signaling through FGFR2c for their survival and expansion. Furthermore, NAMP maintenance depends at least in part on humoral signals distinct from FGF-2. In conclusion, our data show a niche/progenitor organization in vitro, in which the BM adherent fraction provides a self-renewing microenvironment for primitive NAMP.

show abstract

Section: Discussionmentioning

confidence: 73%

“…Since FGF-2 was previously shown to select a subset of MSC with early progenitor characteristics [32], we investigated whether FGF-2 was necessary for NAMP maintenance. Removal of FGF-2 completely abolished the presence of NAMP (Fig.…”

Section: Namp Depend On Fgf-2 Signaling Through Fgfr2cmentioning

confidence: 99%

Fibroblast Growth Factor-2 Maintains a Niche-Dependent Population of Self-Renewing Highly Potent Non-adherent Mesenchymal Progenitors Through FGFR2c

et al. 2012

Self Cite

View full text Add to dashboard Cite

show abstract

“…It is evident from the literature that stem cell expansion conditions can significantly influence their subsequent chondrogenic capacity (Tsutsumi et al 2001;Bianchi et al 2003;Solchaga et al 2005;Khan et al 2008;Solchaga et al 2010). This study develops upon previous work to ascertain if such expansion conditions, specifically expansion in the presence of FGF-2, could further enhance the mechanical functionality of cartilaginous constructs engineered using IFP derived MSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Augmenting MSC expansion conditions to enhance proliferation kinetics while maintaining multipotency is a critical obstacle to overcome in order to engineer mechanically functional tissues for clinical applications. Fibroblast growth factor-2 (FGF-2, also known as basic fibroblast growth factor) has been shown to be a potent stimulator during ex vivo expansion of both chondrocytes (Kato and Gospodarowicz 1985;Martin et al 1999;Martin et al 2001;Veilleux and Spector 2005) and MSCs (Banfi et al 2000;Mastrogiacomo et al 2001;Tsutsumi et al 2001;Bianchi et al 2003;Solchaga et al 2005;Khan et al 2008;Solchaga et al 2010), as well as regulating subsequent differentiation potential. Specifically, bone marrow derived MSCs cultured in the presence of FGF-2 have been shown to be physically smaller and proliferate more rapidly during monolayer expansion (Solchaga et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Buckley

Kelly

2012

Journal of the Mechanical Behavior of Biomedical Materials

View full text Add to dashboard Cite

A bstractMSCs from non-cartilaginous knee joint tissues such as the infrapatellar fat pad (IFP) and synovium possess significant chondrogenic potential and provide a readily available and clinically feasible source of chondroprogenitor cells. Fibroblast growth factor-2 (FGF-2) has been shown to be a potent mitotic stimulator during ex vivo expansion of MSCs, as well as regulating their subsequent differentiation potential.The objective of this study was to investigate the longer term effects of FGF-2 expansion on the functional development of cartilaginous tissues engineered using MSCs derived from the IFP. IFP MSCs were isolated and expanded to passage 2 in a standard media formulation with or without FGF-2 (5ng/ml) supplementation.Expanded cells were encapsulated in agarose hydrogels, maintained in chondrogenic media for 42 days and analysed to determine their mechanical properties and biochemical composition. Culture media, collected at each feed, was also analysed for biochemical constituents.MSCs expanded in the presence of FGF-2 proliferated more rapidly, with higher cell yields and lower population doubling times. FGF-2 expanded MSCs generated the most mechanically functional tissue. Matrix accumulation was dramatically higher after 21 days for FGF-2 expanded MSCs, but decreased between day 21 and 42. By day 42, FGF-2 expanded MSCs had still accumulated ~1.4 fold higher sGAG and ~1.7 fold higher collagen compared to control groups. The total amount of sGAG synthesised (retained in hydrogels and released into the media) was ~ 2.4 fold higher for FGF-2 expanded MSCs, with only ~25% of the total amount generated being retained within the constructs. Further studies are required to investigate whether IFP derived MSCs have a diminished capacity to synthesise other matrix components important in the aggregation, assembly and retention of proteoglycans. In conclusion, expanding MSCs in the presence of FGF-2 rapidly accelerates chondrogenesis in 3D agarose cultures resulting in superior mechanical functionality.

show abstract

“…EGF has also been linked to cell survival in mesenchymal stem cells (Fan et al 2007). Other studies have shown that media containing basic fibroblastic growth factor (bFGF or FGF-2) in MSC culture maintain telomere length and result in a prolonged retention of differentiation potential (Bianchi et al 2003;Yanada et al 2006). The osteogenic and proliferative potential of ASCs can also be preserved by expanding the cells in the presence of bFGF (Quarto and Longaker 2006).…”

Section: Introductionmentioning

confidence: 99%

Monolayer cell expansion conditions affect the chondrogenic potential of adipose‐derived stem cells

Estes

Diekman

Guilak

2008

Biotech & Bioengineering

View full text Add to dashboard Cite

Adipose-derived stem cells (ASCs) are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue. Isolated ASCs are typically expanded in monolayer on standard tissue culture plastic with a basal medium containing 10% fetal bovine serum. However, recent data suggests that altering the monolayer expansion conditions by using suspension culture plastic, adding growth factors to the medium, or adjusting the seeding density may affect the self-renewal rate, multipotency, and lineage-specific differentiation potential of the ASCs. We hypothesized that variation in any of these expansion conditions would influence the chondrogenic potential of ASCs. ASCs were isolated from human liposuction waste tissue and expanded through two passages with different tissue culture plastic, feed medium, and cell seeding densities. Once expanded, the cells were cast in an agarose gel and subjected to identical chondrogenic culture conditions for 7 days, at which point cell viability, radiolabel incorporation, and gene expression were measured. High rates of matrix synthesis upon chondrogenic induction were mostly associated with smaller cells, as indicated by cell width and area on tissue culture plastic, and it appears that expansion in a growth factor supplemented medium is important in maintaining this morphology. All end-point measures were highly dependent on the specific monolayer culture conditions. These results support the hypothesis that monolayer culture conditions may "prime" the cells or predispose them towards a specific phenotype and thus underscore the importance of early culture conditions in determining the growth and differentiation potential of ASCs.

show abstract

Ex vivo enrichment of mesenchymal cell progenitors by fibroblast growth factor 2

Cited by 337 publications

References 42 publications

Fibroblast Growth Factor-2 Maintains a Niche-Dependent Population of Self-Renewing Highly Potent Non-adherent Mesenchymal Progenitors Through FGFR2c

Fibroblast Growth Factor-2 Maintains a Niche-Dependent Population of Self-Renewing Highly Potent Non-adherent Mesenchymal Progenitors Through FGFR2c

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Monolayer cell expansion conditions affect the chondrogenic potential of adipose‐derived stem cells

Contact Info

Product

Resources

About