Kalmaz G D, McDonald T P
Proc Soc Exp Biol Med. 1985 Oct;180(1):50-6. doi: 10.3181/00379727-180-42142.
Plasma clots containing fetal mouse liver cells (FMLC) were used to study the effects of a thrombocytopoiesis-stimulating factor (TSF) from kidney cell culture medium on the proliferation and maturation of megakarocytes. Cells in the megakaryocytic series were identified by the presence of acetylcholinesterase (AChE) and by their morphological and ultrastructural characteristics. For these experiments, 1 X 10(3) to 1 X 10(5) FMLC were cultured for 1-7 days with 0-5 micrograms of TSF; control cultures were treated with production medium (PMC) in which kidney cells had not been grown. The number of AChE+ cells that were observed depended upon the number of cells plated, i.e., after 6 days of culture with 5 micrograms of TSF, an average of 187 AChE+ cells was found after plating 1 X 10(4) cells and 1020 AChE+ cells were observed after plating 1 X 10(5) cells. In dose-response experiments, the number of AChE+ cells rose with increasing doses of TSF. Significantly elevated numbers of AChE+ cells were observed after the addition of 1-5 micrograms of TSF. The optimum time of culture, based upon the number of AChE+ cells found, was 3-5 days. Ultrastructural analysis of megakaryocytes in plasma clots showed evidence of platelet shedding on Day 5. After the culture of FMLC with TSF, a larger number of AChE+ cells was formed from a given number of cells plated than in previous studies that used adult bone marrow cells. Therefore, because of its greater sensitivity, FMLC may be useful for the assay of low levels of TSF, and may be a valuable tool for studying the effects of megakaryocytic regulatory factors on megakaryocytopoiesis.
含有胎鼠肝细胞（FMLC）的血浆凝块被用于研究肾细胞培养基中的血小板生成刺激因子（TSF）对巨核细胞增殖和成熟的影响。巨核细胞系列中的细胞通过乙酰胆碱酯酶（AChE）的存在以及其形态和超微结构特征来鉴定。在这些实验中，将1×10³至1×10⁵个FMLC与0至5微克TSF培养1至7天；对照培养物用未培养肾细胞的生产培养基（PMC）处理。观察到的AChE⁺细胞数量取决于接种的细胞数量，即在用5微克TSF培养6天后，接种1×10⁴个细胞后平均发现187个AChE⁺细胞，接种1×10⁵个细胞后观察到1020个AChE⁺细胞。在剂量反应实验中，AChE⁺细胞数量随TSF剂量增加而增加。添加1至5微克TSF后观察到AChE⁺细胞数量显著增加。根据发现的AChE⁺细胞数量，最佳培养时间为3至5天。血浆凝块中巨核细胞的超微结构分析显示在第5天有血小板脱落的迹象。用TSF培养FMLC后，与之前使用成年骨髓细胞的研究相比，从给定数量的接种细胞中形成了更多的AChE⁺细胞。因此，由于其更高的敏感性，FMLC可能有助于检测低水平的TSF，并且可能是研究巨核细胞调节因子对巨核细胞生成影响的有价值工具。
