Abstract
BackgroundThere is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.ResultsGoat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.ConclusionsOur findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.Electronic supplementary materialThe online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.
Highlights
There is considerable interest in using goats as models for genetically engineering dairy animals and for using stem cells as therapeutics for bone and cartilage repair
At Passage 5 (P5), there were significant differences in Colony-forming efficiency (CFE) between the cell lines (P = 0.0008). 9004 Bone marrow-derived Mesenchymal stem cells (MSCs) (BM-MSCs) exhibited a CFE of 61.0% ± 2.2, which was significantly higher (P < 0.05) than those of 9003 adipose-derived MSCs (ASCs) (48.0 ± 4.2) and 9003 BM-MSC (40.7 ± 1.9) [Figure 1A]
passage 15 (P15) MSCs exhibited a CFE of 47.4% ± 3.4, while passage 10 (P10) MSCs yielded a CFE of 43.8% ± 2.7, which was significantly lower compared to passage 20 (P20) MSCs (P < 0.05) [Figure 1B]
Summary
There is considerable interest in using goats as models for genetically engineering dairy animals and for using stem cells as therapeutics for bone and cartilage repair. Goats are widely used as large animal models for bone tissue engineering as they have knee joints that are similar to humans [12]. Due to the osteogenic and chondrogenic potential of MSCs, goat MSCs are utilized in tissue engineering applications for bone and cartilage regeneration, such as repairing segmental bone defects [13,14] and restoring articular cartilage [15,16]. Reports on the characterization of goat MSCs are relatively more recent and amount to little compared to MSCs isolated from other more commonly used species such as the human, mouse and pig. Many of the reports utilizing goat MSCs, mostly from bone marrow, for tissue engineering lack characterization information. Little information exists on comparisons between goat MSCs isolated from different source tissues, and on comparisons with fibroblasts, which are morphologically similar to and share characteristics with MSCs [24,25]
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