Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There 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.

Results

Goat 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.

Conclusions

Our 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 material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.     

施用羊粪对多花黑麦草生产性能和氮素生产效率的影响

为了探讨羊粪施用量对多花黑麦草生产性能和氮素生产效率的影响,以3个多花黑麦草品种为材料,采用5种羊粪施用梯度进行田间试验。结果表明:在0~60 t/hm2范围内,随着羊粪施用量的增加,多花黑麦草的分蘖数、株高、产量、粗蛋白含量均能显著提高,但氮素的生产效率逐步降低,且3个品种间的差异不显著(P0.05);证实3个品种均适合利用羊粪种植,在多次刈割条件下,最佳羊粪施用量为45~60 t/hm2(折合纯氮778.5~1038 kg/hm2)。     

山羊GH基因5′侧翼序列多态及生物信息学分析

本试验采用PCR-SSCP方法，分析了黄淮山羊、长江三角洲白山羊以及波尔山羊168只个体GH基因5′侧翼区两个位点的遗传多态性，同时分析了三个群体是否为平衡群体。结果如下：在GH基因的两个位点上均发现了多态性，其中第1对引物出现6种基因型，存在2处突变；第2对引物出现3种基因型，存在3处突变。群体分析表明，第1个位点三个群体都处于非平衡状态，说明该位点有经过人工选择的可能；而第2个位点三个群体则都处于平衡状态。进一步利用生物信息学软件分析GH基因的5′侧翼序列，发现在其多态位点处存在IA-1、PIT-1、Gsh-2、C/EBP等多个分值较高的转录因子结合位点，其中264位T→C的突变导致PIT-1和Gsh-2结合位点的消失，至于这些位点是否会影响到GH基因的表达，进而影响到山羊个体的生长发育还需作进一步的研究。     

松嫩平原人工羊草草地放牧绒山羊牧食行为

选取6只2.5岁体重相近而且健康的内蒙古白绒山羊(羯羊、母羊各3只),置于1hrm2人工羊草草地围栏内放牧以观察春、夏、秋、冬四季放牧山羊的行为节律、牧食行为,并计算每天干物质采食量.结果表明:抽穗期羊优先采食草穗,春季过后穗的残余率仅有4.4％;采食速度表现为夏季＞秋季＞春、冬季(P＜0.05);采食时间占方面时间的比例表现为春、秋＞冬、夏季.每口采食量的季节变化趋势大体为春、夏、冬＞秋季(P＜0.05);夏季羊的日采食量最高,其次为春、秋,冬季采食量最低(P＜0.05);同一季节内羯羊的日采食量始终高于母羊(P＜0.05);春、夏、秋季放牧山羊均有3个进食高峰,分别发生在出牧后约0.5h、午后约0.5h、归牧前2h左右,上午采食量低于下午(P＜0.05).冬季一般上午、下午各一个采食高峰;本试验未发现放牧绒山羊扒食草根的现象,但在卧倒休息或反刍前有扒草、扒土整理休息地的习性,未发现放牧山羊行为节律和采食速度的性别差异.