浓缩血小板血浆对造血干细胞增殖分化作用的研究

Effects of the plasma of pooled concentrated platelets on expansion of hematopoietic stem cells

目的观察浓缩血小板血浆对外周血造血干细胞增殖分化的作用。方法使用淋巴细胞分离液分离外周血单个核细胞后进行培养;按在IMDM培养液中加入血浆的不同分为3组:浓缩血小板保存0d(A组)和5d血浆组(C组)、单采血小板保存5d血浆组(B组);A、B、C组在培养的1、3、7d时取细胞进行瑞氏染色观察细胞形态,流式细胞术检测CD41+、CD61+细胞的增殖情况,C组在培养0、23h时流式细胞术检测CD34+CD38+、CD34+CD38-的增殖情况。结果浓缩血小板保存5d血浆组(C组)培养23h后CD34+CD38-、CD34+CD38+细胞分别扩增了3.12±1.45倍、0.98±0.45倍;浓缩血小板保存0d血浆组(A组)、单采血小板保存5d血浆组(B组)和浓缩血小板保存5d血浆组(C组)培养3d后CD41+细胞分别扩增了1.16±0.36倍、1.28±0.67倍和18.72±3.73倍;CD61+细胞分别扩增了1.67±0.55倍、4.01±1.43倍和19.32±4.32倍。结论浓缩血小板保存5d的血浆具有促进造血干细胞快速增殖的作用,并能在体外诱导巨核系细胞的分化。

Objective To study the effects of the plasma of the allogene pooled concentrated platelets on the expansions of hematopoietic stem cells.Methods The mononuclear cells yielded in apheresis from health donors mobilized by rhGM-CSF were separated by Ficoll density gradient centrifugation and incubated in IMDM medium with 10% human plasma.The study was divided into three groups,group A was added plasma from pooled concentrated platelets stored at d 0,group B was added plasma from apheresis platelets stored at d 5,group C was added plasma from pooled concentrated platelets stored at d 5.The incubated cells were examined by flow cytometer.Results After incubation 23 h,CD3^4＋CD38^-cells were expanded for（3.12±1.45）fold,and CD34^＋CD38^＋cells were expanded for（0.98±0.45）fold.At d 3,CD41^＋ cells in groups A,B and C were expanded（1.16±0.36）,（1.28±0.67）and（18.72±3.73）fold,respectively;CD61^＋ cells were expanded（1.67±0.55）,（4.01±1.43）,（19.32±4.32）fold,respectively.Conclusion The plasma of pooled concentrated platelets can promote the proliferation of hemapotoietic stem cell and accelerate the differentiation of megakaryocyte.