Gene expression data from ahES cells, ES cells, MEF cells and round sperm

Haploid stem cells offer an easy-to-manipulate genetic system and therefore have great values for studies of recessive phenotypes. Here, we show that mouse androgenetic haploid ES (ahES) cell lines can be established by transferring sperm into enucleated oocyte. The ahES cells maintain haploidy and stable growth over 30 passages, express pluripotent markers, possess the ability to differentiate into all three germ-layers in vitro and in vivo, and contribute to germline of chimeras when injected into blastocysts. Although epigenetically distinct from sperm cells, the ahES cells can produce viable and fertile progenies after intracytoplasmic injection into mature oocytes. The oocyte injection procedure can also produce viable transgenic mice from genetically engineered ahES cells. We used microarrays to compare the global programme of gene expression among ahES cells, normal diploid ES cells, MEF cells and round sperm cells and found that gene expression pattern of ahES cells was highly similar with ES cells but was distinct from MEF cells and round sperms. Androgenetic haploid ES cells were FACS sorted to harvest the G0/G1 phase haploid cells. Total RNA were extracted from three ahES cell lines (AH129-5, AH129-N1, AH129-NC1, all 129Sv genetic background), two ES cell lines ( CS1-1, R1, 129Sv background), MEF cells and round sperm and hybridized with Affymetrix GeneChip 430 2.0 array. Data were collected and analyzed to compare their gene expression pattern.

单倍体干细胞（haploid stem cells）具备易于操控的遗传体系，因此在隐性表型研究中具有极高价值。本研究证实，小鼠雄核发育单倍体胚胎干细胞（androgenetic haploid ES, ahES）细胞系可通过将精子注入去核卵母细胞的方式构建。该ahES细胞可维持单倍体状态并在30次传代过程中保持稳定生长，表达多能性标志物，具备在体外及体内分化为三胚层的能力，且当被注入囊胚后可参与嵌合体的生殖系形成。尽管在表观遗传层面与精子细胞存在差异，但ahES细胞在通过胞浆内注射注入成熟卵母细胞后，可产生具有存活能力且可育的子代。此外，通过卵母细胞注射流程，还可利用经过基因工程改造的ahES细胞获得存活的转基因小鼠。我们通过微阵列技术比较了ahES细胞、正常二倍体胚胎干细胞、小鼠胚胎成纤维细胞（mouse embryonic fibroblast, MEF）以及圆形精子细胞的全基因表达谱，发现ahES细胞的基因表达模式与胚胎干细胞高度相似，但与MEF细胞及圆形精子细胞存在显著差异。我们通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）对雄核发育单倍体胚胎干细胞进行分选，以获取G0/G1期的单倍体细胞。我们从3株ahES细胞系（AH129-5、AH129-N1、AH129-NC1，均为129Sv遗传背景）、2株胚胎干细胞系（CS1-1、R1，129Sv遗传背景）、MEF细胞以及圆形精子中提取总RNA，并与Affymetrix GeneChip 430 2.0芯片进行杂交。随后对采集到的数据进行分析，以比较各组样本的基因表达模式。