Yoon Sang Hoon, Kim Dae Hee, Cho Sam, Kim Ki-Jeong
Department of Neurosurgery, Armed Forces Capital Hospital, Seongnam, Korea.
Department of Neurosurgery & Spine Care, Seoul National University Bundang Hospital, Seongnam, Korea.
Korean J Neurotrauma. 2020 Oct 16;16(2):138-146. doi: 10.13004/kjnt.2020.16.e36. eCollection 2020 Oct.
We aimed to determine whether bone marrow-derived mesenchymal stem cells (BDMSCs) effectively attenuate the degeneration of human nucleus pulposus cells (NPCs).
Four NPC lines were obtained from 3 subjects who underwent spinal surgery for cervical disc herniation (n=1) or lumbar disc herniation (n=2). For co-culture wells without contact, BDMSCs and adipose-derived mesenchymal stem cells (ADMSCs) were seeded on tissue culture plates and maintained for 3 days. Senescence-associated β-gal (SA-β-gal) staining was represented as a percentage of the total number of stained cells (%). The cells with intracellular lipid droplets (LDs) were represented as the percentage of the number of cells with LDs. Glycosaminoglycan (GAG) secretion was measured at 450 nm, using a commercial kit, to analyze optical density.
The ratio of cells stained with SA-β-gal to the total number of cells reduced significantly when co-cultured with BDMSCs and ADMSCs (<0.001 vs. <0.001). The proportion of NPCs containing LDs was lower when co-cultured with BDMSCs than with ADMSCs (<0.001). The optical density related to GAG secretion was lower in BDMSCs and ADMSCs co-cultured with NPCs than in the controls (<0.001 vs. <0.001).
SA-β-gal staining showed significant attenuation of degenerative changes in NPCs co-cultured with BDMSCs. Moreover, the unexpected increase in LDs was significantly higher in NPCs co-cultured with ADMSCs than in those co-cultured with BDMSCs. However, GAG secretion was significantly decreased in NPCs co-cultured with MSCs.
我们旨在确定骨髓间充质干细胞(BDMSC)是否能有效减轻人髓核细胞(NPC)的退变。
从3例因颈椎间盘突出症(1例)或腰椎间盘突出症(2例)接受脊柱手术的患者中获取4株NPC系。对于非接触共培养孔,将BDMSC和脂肪间充质干细胞(ADMSC)接种于组织培养板上并培养3天。衰老相关β-半乳糖苷酶(SA-β-gal)染色以染色细胞总数的百分比(%)表示。含有细胞内脂滴(LD)的细胞以有LD的细胞数量的百分比表示。使用商业试剂盒在450nm处测量糖胺聚糖(GAG)分泌,以分析光密度。
与BDMSC和ADMSC共培养时,SA-β-gal染色细胞与细胞总数的比例显著降低(<0.001对<0.001)。与BDMSC共培养时,含有LD的NPC比例低于与ADMSC共培养时(<0.001)。与NPC共培养的BDMSC和ADMSC中与GAG分泌相关的光密度低于对照组(<0.001对<0.001)。
SA-β-gal染色显示与BDMSC共培养的NPC退变变化明显减轻。此外,与BDMSC共培养的NPC中意外出现的LD增加显著高于与ADMSC共培养的NPC。然而,与间充质干细胞共培养的NPC中GAG分泌显著减少。