目的通过观察再生障碍性贫血(再障)患者间充质干细胞(MSCs)成脂肪和成骨能力的变化,研究骨髓MSCs成脂分化的异常在再障患者红髓脂肪化中的作用。方法分离培养再障患者及正常人的骨髓,观察其一般生物学特性,并在体外诱导其向脂肪、成骨细胞分化,同时用RT—PCR方法检测成脂肪、成骨特异基因的表达时间,比较再障患者和正常对照MSCs向脂肪细胞、成骨分化能力的不同。结果原代培养7d,再障组贴壁细胞克隆形成率为(19.30±4.77)/(5×10^5MNCs),较正常对照组明显降低(47.72±3.46)/(5×10^5MSCs)(P〈0.05)。在培养初期,再障组MSCs与正常对照MSCs增殖能力相似,但在连续传8代后,其增殖能力降低。再障组MSCs体外诱导成脂滴早,诱导分化的脂肪细胞leptin(瘦素)基因表达早。再障组MSCs体外诱导成骨形成钙化结节少,碱性磷酸酶活性低,诱导的成骨细胞osteocalcin(骨钙素)基因表达晚。结论再障患者MSCs成脂分化能力增强而成骨分化能力降低,可能在再障的病程中起一定作用。
Objective To investigate the osteogenic and adipogenic difference of bone marrowderived mesenchymal stem cells (MSCs) between patients with aplastic anemia(AA) and healthy volunteers and to explore the role of MSCs adipo-differentiation in the pathogenetic mechanism of AA. Methods MSCs were isolated from bone marrow of patients with AA and healthy donors and expanded in vitro. MSCs derived from the AA patients and healthy volunteers were compared with respect to morphology, in vitro proliferation capacity, phenotype, differentiation ability and gene expression during differentiation. Results The MSCs clones in the AA patients were( 19. 30 ± 4. 77)/(5 ×10^5 MNCs) 7 days after culture, being significantly lower than those in the healthy volunteers, which was (47.72 ±3.46 )/(5 ×10^5MSCs ) ( P 〈 0. 05 ). Compared with those the healthy donors, MSCs from the AA patients had similar proliferative capacity in the first 8 passages and then decreased in the following passages. MSCs from different sources had the same phenotype, MSCs from thc AA patients could differentiate more easily into adipocytes but less easily and slower into osteoblasts than those from the healthy volunteers. Conclusion The increased adipogenic capacity and decreased osteogenic capacity of MSCs in AA patients may contribute to the development and progress of AA.