McDonald T P, Jackson C W
University of Tennessee College of Veterinary Medicine, Knoxville 37901-1071.
Exp Hematol. 1990 Aug;18(7):758-63.
A thrombocytopoiesis-stimulating factor (TSF or thrombopoietin) is known to increase the size and number of mouse bone marrow megakaryocytes, to increase the proportion of megakaryocytes in endomitosis, and to increase the number of small acetylcholinesterase-positive cells. Megakaryocyte ploidy values have not previously been measured in mice treated with TSF from human embryonic kidney (HEK) cells. Therefore, in the present study C3H mice were injected with approximately 4 U of step II TSF; platelet production indices and megakaryocyte ploidy values were measured 1-5 days after treatment. For controls, other mice were injected with saline, human serum albumin (HSA), normal rabbit serum (NRS), or rabbit anti-mouse platelet serum (RAMPS). Platelet counts, platelet sizes, and percent 35S incorporation into platelets were measured using standard techniques. For measurement of megakaryocyte DNA content, bone marrow cells were collected into CATCH medium and incubated with RAMPS, followed by labeling with a saturating concentration of fluorescein-conjugated goat anti-rabbit immunoglobulin F(ab')2 fragment. After washing, the cells were resuspended in propidium iodide, and DNA content was measured by flow cytometry. When compared to suitable control values, the results showed that TSF caused a significant (p less than 0.025) increase in platelet counts of treated mice by 3 days; both TSF and RAMPS caused significant (p less than 0.0005) increases in platelet sizes and percent 35S incorporation into platelets of mice at 2 and 3 days after treatment. The most frequent polyploid DNA class of megakaryocytes from untreated C3H mice was 32N, confirming our previous observation. Both TSF and RAMPS caused significant (p less than 0.0005) increases in the average polyploid megakaryocyte DNA content, with peak values on days 2 and 3. These data show that TSF administered in vivo significantly increases DNA content of mouse bone marrow megakaryocytes.
已知一种血小板生成刺激因子（TSF 或血小板生成素）可增加小鼠骨髓巨核细胞的大小和数量，增加处于核内有丝分裂的巨核细胞比例，并增加小的乙酰胆碱酯酶阳性细胞数量。此前尚未对用来自人胚胎肾（HEK）细胞的TSF处理的小鼠的巨核细胞倍性值进行测量。因此，在本研究中，给C3H小鼠注射约4 U的II期TSF；在处理后1 - 5天测量血小板生成指数和巨核细胞倍性值。作为对照，给其他小鼠注射生理盐水、人血清白蛋白（HSA）、正常兔血清（NRS）或兔抗小鼠血小板血清（RAMPS）。使用标准技术测量血小板计数、血小板大小以及35S掺入血小板的百分比。为了测量巨核细胞DNA含量，将骨髓细胞收集到CATCH培养基中，与RAMPS一起孵育，然后用饱和浓度的荧光素偶联山羊抗兔免疫球蛋白F(ab')2片段进行标记。洗涤后，将细胞重悬于碘化丙啶中，通过流式细胞术测量DNA含量。与合适的对照值相比，结果显示TSF使处理后3天的小鼠血小板计数显著增加（p小于0.025）；TSF和RAMPS均使处理后2天和3天的小鼠血小板大小以及35S掺入血小板的百分比显著增加（p小于0.0005）。未处理的C3H小鼠的巨核细胞中最常见的多倍体DNA类别是32N，证实了我们之前的观察结果。TSF和RAMPS均使平均多倍体巨核细胞DNA含量显著增加（p小于0.0005），在第2天和第3天达到峰值。这些数据表明，体内给予TSF可显著增加小鼠骨髓巨核细胞的DNA含量。
