真性红细胞增多症患者骨髓间充质干细胞体外造血支持功能研究

Study on hematopoietic supportive properties of bone marrow stem cell of patients with polycythemia vera

目的研究真性红细胞增多症(PV)患者骨髓间充质干细胞(BMSC)的造血支持功能。方法从PV初诊患者和健康志愿者骨髓单个核细胞中分离培养BMSC;光学显微镜观察细胞形态;流式细胞术鉴定细胞免疫表型;诱导成骨及成脂分化鉴定细胞多向分化能力;与人脐带血CD34+造血干祖细胞(HSPC)共培养检测真性红细胞增多症患者来源的骨髓间充质干细胞(PV-BMSC)的体外造血支持功能。结果 PV-BMSC的细胞形态、免疫表型与正常人来源的骨髓间充质干细胞(NM-BMSC)相比无明显差异;油红O及Von Kossa染色表明PV-BMSC具有成脂和成骨分化能力;实验共分离得9株PV-BMSC,其中55%的PV-BMSC既能支持总细胞增殖,又能支持CD34+HSPC扩增(≥1.5倍,n=5),22%的仅支持总细胞增殖(≥1.5倍,n=2),其余的既不支持总细胞增殖也不支持CD34+HSPC扩增(≤1.0倍,n=2)。结论 PV-BMSC支持CD34+HSPC增殖存在明显的个体差异。

Objective To study the hematopoietic supportive properties of bone marrow stem cell （BMSC） of patients with polycythemia vera （PV）. Methods BMSC were isolated and cultured from the bone marrow mononuclear celis of newly diagnosed PV patients arLd healthy volunteers. The cell morphology, immunophenotype and multi-directional differentiation of PV-BMSC were identified by optical microscope, flow cytometry as well as adipogenic and os- teogenic differentiation culture, respectively. The hematopoietic supportive property in vitro of BMSC from patients with PV （PV-BMSC） was evaluated via co-culturing with CD34＋ hematopoietic stem and progenitor ceils （HSPC） de- rived from human umbilical-coat blood. Results The isolated PV-BMSC shared similar cellular morphology and sur- face marker expression profile with BMSC from normal bone marrow （NM-BMSC）. Oil Red O and Von Kossa staining indicated that PV-BMSC had multi-lineage differentiation potential. Among the isolated 9 PV-BMSC cell lines, 55% of the PV-BMSC supported both cell proliferation and CD34＋ HSPC expansion （≥ 1.5 folds, n = 5）, 22% only sup- ported cell proliferation （≥ 1.5 iblds, n = 2） and 22% supported neither cell proliferation nor CD34＋ HSPC expansion（〈 1.0 fold, n = 2）. Conclusion PV-BMSC show sig- nificant variability in supporting CD34 ＋ cell expansion.