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豚鼠分离巨核细胞的体外培养:恢复、存活、形态学变化及成熟

Culture in vitro of isolated guinea pig megakaryocytes: recovery, survival, morphologic changes, and maturation.

作者信息

Levine R F

出版信息

Blood. 1977 Oct;50(4):713-25.

PMID:901941
Abstract

Isolated guinea pig megakaryocytes were maintained in liquid cultures for up to 4 days. Megakaryocytes were incubated in siliconized glass vials in Dulbecco's Modified Eagle Medium with 5%-10% guinea pig serum and 2.3% bovine serum albumin. Cultured megakaryocytes did not adhere to glass vials and were almost entirely recovered by aspiration. No reproduction or cell division of megakaryocytes occurred. A small decline in viability occurred promptly on placing the freshly isolated cells in culture medium and could be attributed to reexposure to calcium. On incubation there was little further cell death. Up to 2 days in culture the megakaryocytes remained morphologically intact and appeared similar to megakaryocytes in situ. Megakaryocytes matured in culture with a loss of cytoplasmic basophilia, an increase in granule content, and progressive changes in nuclear configuration. The most mature megakaryocytes developed pseudopod formation but large-scale platelet liberation was not seen. The ability to culture megakaryocytes in vitro will allow more extensive biochemical and physiologic studies of this cell than previously possible.

摘要

将分离出的豚鼠巨核细胞在液体培养中维持长达4天。巨核细胞在硅化玻璃小瓶中,于含有5%-10%豚鼠血清和2.3%牛血清白蛋白的杜尔贝科改良伊格尔培养基中孵育。培养的巨核细胞不粘附于玻璃小瓶,几乎可通过抽吸完全回收。巨核细胞未发生增殖或细胞分裂。将刚分离的细胞置于培养基中后,活力立即出现轻微下降,这可能归因于再次暴露于钙。孵育时几乎没有进一步的细胞死亡。培养2天内,巨核细胞在形态上保持完整,与原位巨核细胞相似。巨核细胞在培养中成熟,胞质嗜碱性降低,颗粒含量增加,核构型逐渐改变。最成熟的巨核细胞形成伪足,但未见大规模血小板释放。与以前相比,体外培养巨核细胞的能力将使对这种细胞进行更广泛的生化和生理学研究成为可能。

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