非接触式共培养下两种同源间充质干细胞对退变髓核细胞生物学影响的比较研究

Comparison of the biological effects of two kinds of homologous mesenchymal stem cells on degenerative nucleus pulposus cells under the condition of non-contact co-culture

中文关键词:  非接触式共培养  同源间充质干细胞  退变髓核细胞  比较研究

英文关键词:Non-contact co-culture  Homologous mesenchymal stem cells  Degenerative nucleus pulposus cells  Ccomparative study

中文摘要:

  【摘要】 目的：比较非接触式共培养条件下两种同源间充质干细胞对退变髓核细胞生物学功能的影响。方法：取同一腰椎间盘突出症患者骨髓间充质干细胞（BMSCs）、脂肪间充质干细胞（ADSCs）和退变髓核细胞（NPCs），两种间充质干细胞分别与髓核细胞在Tanswell 6孔板中进行非接触式共培养，与ADSCs共培养的NPCs为A组，与BMSCs共培养的NPCs为B组，以单独培养的NPCs为对照组。共培养7d后，提取3组髓核细胞总RNA，进行反转录后，利用Real-Time PCR检测其Ⅱ型胶原、蛋白多糖和SOX-9基因的相对表达量。结果：非接触式共培养7d后，Ⅱ型胶原、蛋白多糖和SOX-9基因相对表达量对照组分别是1.03±0.28、1.21±0.40和0.94±0.34，A组分别是3.49±0.55、3.88±2.11和2.41±0.91，B组分别是7.60±1.89、6.26±2.96和4.55±1.88。与对照组相比，A、B组髓核细胞Ⅱ型胶原、蛋白多糖和SOX-9基因相对表达显著增加（P<0.05）；A、B两组间相比也有显著性差异（P<0.05）。结论：非接触式共培养条件下骨髓间充质干细胞和脂肪间充质干细胞对退变髓核细胞均有一定的激活效应；骨髓间充质干细胞对退变髓核细胞的激活效应更强，可能更加适合于椎间盘退行性疾病的生物学治疗。

英文摘要:

  【Abstract】 Objectives: To compare the biological effects of degenerative nucleus pulposus cells when co-culturing two kinds of people′s mesenchymal stem cells under a non-contact system. Methods: Bone marrow mesenchymal stem cells(BMSCs), adipose mesenchymal stem cells(ADSCs) and nucleus pulposus cells(NPCs) were obtained from the same patient, then the two kinds of stem cells were co-cultured respectively with degenerative nucleus pulposus cells in Transwell six-well plates under the non-contact condition, NPCs co-cultivation with ADSCs as group A, and NPCs co-cultivation with BMSCs as group B, nucleus pulposus cells cultured alone as control. One week later, the total RNA of 3 groups were extracted, then the Real-Time PCR technology was used to detect the relative gene expression of type Ⅱ collagen, proteoglycan and SOX-9. Results: The relative gene expression of type Ⅱ collagen protein, proteoglycan and SOX-9 in control group was 1.03±0.28, 1.21±0.40 and 0.94±0.34 respectively, in group A was 3.49±0.55, 3.88±2.11 and 2.41±0.91 respectively, while 7.60±1.89, 6.26±2.96 and 4.55±1.88 respectively in group B, The Real-Time PCR results showed that compared with the control group, the nucleus pulposus cells of group A and B presented a significant increase of type Ⅱ collagen protein, proteoglycan and SOX-9 gene expression(P<0.05), which also showed significant difference between groups A and B(P<0.05). Conclusions: Bone marrow mesenchymal stem cells and adipose mesenchymal stem cells can both stimulate and activate the degenerative nucleus pulposus cells under the condition of noncontact co-culture; with stronger effect on the former, which may act as a better option for biological treatment for degenerative disc diseases.