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Combined influence of basal media and fibroblast growth factor on the expansion and differentiation capabilities of adipose-derived stem cells.

Ahearne M, Lysaght J, Lynch AP - Cell Regen (Lond) (2014)

Bottom Line: FGF has a negative effective on passage 4 ASC adipogenesis and high glucose media plus FGF-enhanced osteogenic capacity of passage 4 ASCs.High glucose media plus FGF appeared to be the most beneficial for priming ASCs to induce a keratocyte phenotype.These findings demonstrate the reciprocal effect FGF and basal media have on ASCs.

View Article: PubMed Central - PubMed

Affiliation: Trinity Centre for Bioengineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland ; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Dublin, Ireland.

ABSTRACT

Background: Interest in adipose-derived stem cells (ASCs) has increased in recent years due to their multi-linage differentiation capabilities. While much work has been done to optimize the differentiation media, few studies have focused on examining the influence of different expansion media on cell behavior. In this study, three basal media (low glucose Dulbecco's modified Eagle's medium (DMEM), high glucose DMEM and DMEM-F12) supplemented with or without fibroblast growth factor 2 (FGF) were examined to assess their suitability for expanding ASCs.

Findings: Flow cytometry, colony-forming unit assays (CFU-Fs) and differentiation assays were utilized to study cell behavior. High glucose media CFU-Fs produced fewest colonies while the addition of FGF increased colony size. By passage 2, the majority of cells were positive for CD44, 45, 73, 90 and 105 and negative for CD14, 31 and 45, indicating a mesenchymal phenotype. A sub-population of CD34 positive cells was present among passage 2 cells; however, by passage 4 the cells were negative for CD34. FGF has a negative effective on passage 4 ASC adipogenesis and high glucose media plus FGF-enhanced osteogenic capacity of passage 4 ASCs. FGF supplemented basal media were most suitable for chondrogenesis. High glucose media plus FGF appeared to be the most beneficial for priming ASCs to induce a keratocyte phenotype.

Conclusions: These findings demonstrate the reciprocal effect FGF and basal media have on ASCs. This research has implications for those interested regenerating bone, cartilage, cornea or adipose tissues.

No MeSH data available.


Related in: MedlinePlus

CFU-F assays showing the number of colonies formed after 14 days from freshly isolated cells from stromal vascular fraction of adipose tissue cultured under different media conditions.n = 3 dishes counted per samples, *significant difference with all other groups, **significant different with HG, F12, LG + FGF and HG + FGF, p < 0.05.
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Fig2: CFU-F assays showing the number of colonies formed after 14 days from freshly isolated cells from stromal vascular fraction of adipose tissue cultured under different media conditions.n = 3 dishes counted per samples, *significant difference with all other groups, **significant different with HG, F12, LG + FGF and HG + FGF, p < 0.05.

Mentions: The number and size of colonies formed from freshly isolated ASCs in CFU-Fs varied considerably when using different media types (Figure 2). F12 and LG produced significantly more colonies than HG media. Supplementation with FGF resulted in fewer colonies in F12 and LG media, but much larger colonies were formed. Normally, increasing the number of colonies would suggest a higher proportion of stem cells among the freshly isolated cell population. However, since the same initial number of cells was used in all groups, this demonstrates that the culture conditions also affect colony formation and expansion.Figure 1


Combined influence of basal media and fibroblast growth factor on the expansion and differentiation capabilities of adipose-derived stem cells.

Ahearne M, Lysaght J, Lynch AP - Cell Regen (Lond) (2014)

CFU-F assays showing the number of colonies formed after 14 days from freshly isolated cells from stromal vascular fraction of adipose tissue cultured under different media conditions.n = 3 dishes counted per samples, *significant difference with all other groups, **significant different with HG, F12, LG + FGF and HG + FGF, p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4325938&req=5

Fig2: CFU-F assays showing the number of colonies formed after 14 days from freshly isolated cells from stromal vascular fraction of adipose tissue cultured under different media conditions.n = 3 dishes counted per samples, *significant difference with all other groups, **significant different with HG, F12, LG + FGF and HG + FGF, p < 0.05.
Mentions: The number and size of colonies formed from freshly isolated ASCs in CFU-Fs varied considerably when using different media types (Figure 2). F12 and LG produced significantly more colonies than HG media. Supplementation with FGF resulted in fewer colonies in F12 and LG media, but much larger colonies were formed. Normally, increasing the number of colonies would suggest a higher proportion of stem cells among the freshly isolated cell population. However, since the same initial number of cells was used in all groups, this demonstrates that the culture conditions also affect colony formation and expansion.Figure 1

Bottom Line: FGF has a negative effective on passage 4 ASC adipogenesis and high glucose media plus FGF-enhanced osteogenic capacity of passage 4 ASCs.High glucose media plus FGF appeared to be the most beneficial for priming ASCs to induce a keratocyte phenotype.These findings demonstrate the reciprocal effect FGF and basal media have on ASCs.

View Article: PubMed Central - PubMed

Affiliation: Trinity Centre for Bioengineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland ; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Dublin, Ireland.

ABSTRACT

Background: Interest in adipose-derived stem cells (ASCs) has increased in recent years due to their multi-linage differentiation capabilities. While much work has been done to optimize the differentiation media, few studies have focused on examining the influence of different expansion media on cell behavior. In this study, three basal media (low glucose Dulbecco's modified Eagle's medium (DMEM), high glucose DMEM and DMEM-F12) supplemented with or without fibroblast growth factor 2 (FGF) were examined to assess their suitability for expanding ASCs.

Findings: Flow cytometry, colony-forming unit assays (CFU-Fs) and differentiation assays were utilized to study cell behavior. High glucose media CFU-Fs produced fewest colonies while the addition of FGF increased colony size. By passage 2, the majority of cells were positive for CD44, 45, 73, 90 and 105 and negative for CD14, 31 and 45, indicating a mesenchymal phenotype. A sub-population of CD34 positive cells was present among passage 2 cells; however, by passage 4 the cells were negative for CD34. FGF has a negative effective on passage 4 ASC adipogenesis and high glucose media plus FGF-enhanced osteogenic capacity of passage 4 ASCs. FGF supplemented basal media were most suitable for chondrogenesis. High glucose media plus FGF appeared to be the most beneficial for priming ASCs to induce a keratocyte phenotype.

Conclusions: These findings demonstrate the reciprocal effect FGF and basal media have on ASCs. This research has implications for those interested regenerating bone, cartilage, cornea or adipose tissues.

No MeSH data available.


Related in: MedlinePlus