Talbot Neil C, Caperna Thomas J, Powell Anne M, Ealy Alan D, Blomberg Le Ann, Garrett Wesley M
Agricultural Research Service, Animal and Natural Resources Institute, Biotechnology and Germplasm Laboratory, U.S. Department of Agriculture, Beltsville Agricultural Research Center, Beltsville, MD 20705, USA.
In Vitro Cell Dev Biol Anim. 2005 May-Jun;41(5-6):130-41. doi: 10.1290/040901.1.
A cell line, BPE-1, was derived from a parthenogenetic 8-d in vitro-produced bovine blastocyst that produced a cell outgrowth on STO feeder cells. The BPE-1 cells resembled visceral endoderm previously cultured from blastocysts produced by in vitro fertilization (IVF). Analysis of the BPE-1 cells demonstrated that they produced serum proteins and were negative for interferon-tau production (a marker of trophectoderm). Transmission electron microscopy revealed that the cells were a polarized epithelium connected by complex junctions resembling tight junctions in conjunction with desmosomes. Rough endoplasmic reticulum was prominent within the cells as were lipid vacuoles. Immunocytochemistry indicated the BPE-1 cells had robust microtubule networks. These cells have been grown for over 2 yr for multiple passages at 1:10 or 1:20 split ratios on STO feeder cells. The BPE-1 cell line presumably arose from embryonic cells that became diploid soon after parthenogenetic activation and development of the early embryo. However, metaphase spreads prepared at passage 41 indicated that the cell population had a hypodiploid (2n = 60) unimodal chromosome content with a mode of 53 and a median and mean of 52. The cell line will be of interest for functional comparisons with bovine endoderm cell lines derived from IVF and nuclear transfer embryos.
细胞系BPE-1源自孤雌生殖的体外培养8天的牛囊胚,该囊胚在STO饲养细胞上产生细胞生长物。BPE-1细胞类似于先前从体外受精(IVF)产生的囊胚中培养的脏内胚层。对BPE-1细胞的分析表明,它们能产生血清蛋白,且不产生干扰素τ(滋养外胚层的标志物)。透射电子显微镜显示,这些细胞是极化上皮细胞,通过类似于紧密连接并伴有桥粒的复杂连接相连。细胞内粗面内质网和脂质空泡都很突出。免疫细胞化学表明BPE-1细胞具有强大的微管网络。这些细胞已在STO饲养细胞上以1:10或1:20的分裂比例传代培养超过2年。BPE-1细胞系可能源自孤雌生殖激活和早期胚胎发育后不久变成二倍体的胚胎细胞。然而,在第41代制备的中期染色体铺展显示,细胞群体具有亚二倍体(2n = 60)单峰染色体含量,众数为53,中位数和平均数为52。该细胞系对于与源自IVF和核移植胚胎的牛内胚层细胞系进行功能比较具有重要意义。