Immunomagnetic bead isolation of megakaryocytes from guinea-pig bone marrow: effect of recombinant interleukin-6 on size, ploidy and cytoplasmic fragmentation.
Immunomagnetic bead isolation of megakaryocytes from guinea-pig bone marrow: effect of recombinant interleukin-6 on size, ploidy and cytoplasmic fragmentation.
复制标题
豚鼠骨髓中巨核细胞的免疫磁珠分离：重组白细胞介素 6 对大小、倍性和细胞质碎片的影响。
DOI：
10.1111/j.1365-2141.1991.tb08569.x
复制
发表时间：
1991
影响因子：
6.5
通讯作者：
Rodriguez,A
中科院分区：
文献类型：
--
作者：
Leven,RM;Rodriguez,A
使用血小板糖蛋白Ib的抗体和与磁珠偶联的第二抗体分离豚鼠骨髓巨核细胞。该程序从两只豚鼠中平均获得644800个巨核细胞，平均活力为83%。所有血小板糖蛋白Ib阳性细胞也表达血小板糖蛋白IIb-IIIa复合物。在存在10 ng/ml白细胞介素-6（IL-6）的情况下，分析通过该技术分离的巨核细胞的大小和倍性。在没有IL-6的情况下，巨核细胞的大小在24小时后显著增加。但在IL-6存在下，尺寸甚至更大地增加。典型倍性类型为16 N，平均19%为2 N。4N为2.6%，8N为16.4%。通过流式细胞术测定，50·8%的16N和11·1%的32N细胞。显微分光光度法的测量结果非常一致。培养24小时后，2N和32N细胞的百分比显著增加。用IL-6处理24小时后的倍性分布与对照相同。在不存在血清的情况下，在胶原凝胶上培养的巨核细胞不会形成伪足和碎片，而血清会发生伪足和碎片（Levenet al.1987）。向无血清培养物中添加IL-6导致巨核细胞形成广泛的前血小板延伸。我们的结论是，大量的纯豚鼠骨髓巨核细胞可以通过免疫磁珠选择，包括低倍性的未成熟的巨核细胞分离。巨核细胞大小和倍性的增加证明了自发成熟的发生。IL-6改变巨核细胞的大小和形态，但不改变倍性，表明巨核细胞的这些不同特征可能是单独调节的。
Guinea‐pig bone marrow megakaryocytes were isolated using an antibody to platelet glycoprotein Ib and a second antibody conjugated to magnetic beads. The procedure yielded an average of 644 800 megakaryocytes from two guinea‐pigs with an average viability of 83%. All of the platelet glycoprotein Ib positive cells also expressed the platelet glycoprotein IIb–IIIa complex. The size and ploidy of megakaryocytes isolated by this technique were analysed in the presence of 10 ng/ml of interleukin‐6 (IL‐6). Without IL‐6 megakaryocyte size increased significantly after 24 h. but an even larger increase in size occurred in the presence of IL‐6. The modal ploidy class was 16N with an average of 19% 2N. 2·6% 4N, 16·4% 8N. 50·8% 16N and 11·1% 32N cells as determined by flow cytometry. Measurements made by microspectrophotometry were in close agreement. After 24 h incubation there was a significant rise in the percentage of 2N and 32N cells. The ploidy distribution after 24 h with IL‐6 was the same as the control. Megakaryocytes cultured in the absence of serum on collagen gels did not form pseudopods and fragment, as occurs with serum (Levenet al.1987). Addition of IL‐6 to the serum‐free cultures caused megakaryocytes to form extensive proplatelet extensions. We conclude that large numbers of pure guinea‐pig bone marrow megakaryocytes can be isolated by immunomagnetic bead selection, including low ploidy immature megakaryocytes. Spontaneous maturation occurred as evidenced by the increase in megakaryocyte size and ploidy. IL‐6 altered megakaryocyte size and morphology but not ploidy, indicating that these different characteristics of megakaryocytes may be regulated separately.