Talbot N C, Worku M, Paape M J, Grier P, Rexroad C E, Pursel V G
USDA, LPSI, ARS, Gene Evaluation and Mapping Laboratory, Beltsville, Maryland 20705, USA.
In Vitro Cell Dev Biol Anim. 1996 Oct;32(9):541-9. doi: 10.1007/BF02722981.
Secondary macrophage cell cultures were generated from the primary culture of epiblasts of 8-d-old pig blastocysts. The epiblast-derived macrophagelike (EDM) cells have a morphology and ameboid behavior that is typical of tissue histocytes. The cells reacted positively with monoclonal antibodies specific for pig granulocyte-macrophage lineage cells, and were not reactive with monoclonal antibodies specific for pig B and T lymphocytes. Marked phagocytic behavior and the formation of phagosomes were demonstrated following incubation with FITC-labeled bacteria. The EDM cells stained positively for nonspecific acid esterase that was not inhibited by sodium fluoride. DiI-acetylated-LDL was rapidly taken up by the cells. Transmission electron microscopy of the EDM cells showed phagolysosomes, numerous cytoplasmic vacuoles, large, lobed nuclei, and numerous pseudopods or filopodia at the cell surface. Strong reactivity of the cells with anti-CD14 monoclonal antibody was observed. Further, cytotoxic activity was produced from the EDM cells after exposure to lipopolysaccharide in a concentration and time-dependent manner. The cultures could be maintained and expanded for several months on STO co-culture. Their derivation from the epiblast of the pig demonstrates the possibility of obtaining hemopoietic cell cultures from the preimplantation blastocysts of all mammals.
次级巨噬细胞培养物源自8日龄猪囊胚上胚层的原代培养。上胚层来源的巨噬样(EDM)细胞具有组织组织细胞典型的形态和阿米巴样行为。这些细胞与猪粒细胞-巨噬细胞系细胞特异性单克隆抗体呈阳性反应,而与猪B和T淋巴细胞特异性单克隆抗体无反应。与异硫氰酸荧光素标记的细菌孵育后,显示出明显的吞噬行为和吞噬体形成。EDM细胞对不受氟化钠抑制的非特异性酸性酯酶呈阳性染色。二碘乙酰化低密度脂蛋白被细胞迅速摄取。EDM细胞的透射电子显微镜显示有吞噬溶酶体、大量细胞质空泡、大的分叶核以及细胞表面大量的伪足或丝状伪足。观察到细胞与抗CD14单克隆抗体有强烈反应。此外,EDM细胞在暴露于脂多糖后以浓度和时间依赖性方式产生细胞毒性活性。这些培养物在STO共培养中可维持和扩增数月。它们源自猪的上胚层,这表明从所有哺乳动物的植入前囊胚中获得造血细胞培养物是可能的。