Interest in adipose-derived stem cells (ASCs) has increased in recent years due to their multi-linag

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* Corresponding author: Mark Ahearne ahearnm@tcd.ie
The electronic version of this article is the complete one and can be found online at: http://www.cellregenerationjournal.com/content/3/1/13 Received: 11 July 2014 Accepted: 18 November 2014 Published: 26 November 2014
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication ian mackenzie jeffers waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Interest in adipose-derived stem cells (ASCs) has increased in recent years due to their multi-linage differentiation capabilities. ian mackenzie jeffers 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 ian mackenzie jeffers 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 ian mackenzie jeffers size. By passage 2, the majority of cells were positive for CD44, 45, 73, 90 and 105 and negative ian mackenzie jeffers for CD14, 31 and 45, indicating ian mackenzie jeffers a mesenchymal phenotype. A sub-population ian mackenzie jeffers 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 ian mackenzie jeffers 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. Keywords: Cell differentiation; Cartilage; Bone; Adipose; Cornea Findings
The ability of adipose-derived stem cells (ASCs) to undergo prolonged periods of expansion and differentiation ian mackenzie jeffers towards specific lineages has made them a valuable cell source for clinicians and researchers interested in tissue repair and regenerative medicine. These cells are considered to exhibit mesenchymal stem cell characteristics by their ability to form bone, cartilage and adipose tissues and by the presence or absence of specific CD markers [ 1 ]. Maintaining a suitable culture environment is essential for retaining the stem-like characteristics of ASCs. Commercially available media have been formulated to cultivate these cells; however, these are generally more expensive than standard culture media and their effectiveness over standard chemically defined supplemented media has been brought into question [ 2 ]. For these reasons, many researchers continue to use media consisting of Dulbecco s modified Eagle s medium (DMEM) supplemented with fetal bovine serum (FBS), antibiotics and in some cases growth factors most commonly fibroblast growth factor 2 (FGF). Low glucose DMEM [ 3 , 4 ], high glucose DMEM [ 5 , 6 ] and DMEM plus nutrient mix F12 [ 7 , 8 ] are among the most commonly used basal media for expanding ASCs, although many publications do not state the media glucose concentration. There appears ian mackenzie jeffers to be little consensus ian mackenzie jeffers as to which of these media sources and growth factors are most suitable for ASC expansion. The aims of this study were to compare three of the most commonly used basal media for ASC expansion (low glucose DMEM, high glucose DMEM and DMEM-F12) and determine whether they had any advantages or limitations. The influence of adding FGF to each of these media was also examined. After expansion in each type of media, the cells were placed in chemically defined differentiation media to promote an adipogenic, osteogenic, chondrogenic or keratocyte phenotype. ASC differentiation was evaluated and used to determine the most effective expansion ian mackenzie jeffers media for each specific lineage. Materials and methods
Subcutaneous adipose tissue was taken with consent from patients undergoing esophageal surgery. This study has received ethical approval from the St. James s Hospital Ins