中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2012年
5期
438-442
,共5页
脐带%间充质干细胞%神经营养困子
臍帶%間充質榦細胞%神經營養睏子
제대%간충질간세포%신경영양곤자
Umbilical cord%Mesenchymal stromal cell%Neurotrophic factor
目的 通过检测人脐带间充质干细胞(hUC-MSCs)的神经营养因子谱,以进一步认识hUC-MSCs通过分泌神经营养因子治疗神经损伤的作用. 方法 组织块培养法分离培养hUC-MSCs,流式细胞仪鉴定干细胞表面标记物,Raybio人细胞因子抗体芯片检测hUC-MSCs神经营养因子表达情况,RT-PCR检测hUC-MSCs神经营养因子的基因表达,ELISA法定量检测hUC-MSCs上清液中分泌的脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)、肝细胞生长因子(HGF)、神经营养因子3(NT-3)含量. 结果 流式细胞分析结果显示hUC-MSCs表面标记物CD29、CD44、CD90阳性(94.05％、90.75％、98.12％),CD34、CD45阴性(3.09％、0.80％).Raybio人细胞因子抗体芯片结果显示,与神经营养相关的因子BDNF、表皮生长因子(EGF)、GDNF、HGF、NT-3、血管内皮细胞生长因子(VEGF)、胰岛素样生长因子-1(IGF-1)、神经营养因子4(NT-4)在hUC-MSCs中均有表达.RT-PCR检验也证实IGF-1、VEGF、神经生长因子(NGF)、HGF、GDNF、NT-4、BDNF和NT-3基因在hUC-MSCs表达.ELISA检测细胞上清液中BDNF、GDNF、HGF、NT-3的含量分别为(475.20±32.22) pg/mL、(82.33±3.39) pg/mL、(704.50±12.86) pg/mL、(230.41±16.66) pg/mL,分别与空白对照组相比,差异均有统计学意义(P＜0.05). 结论 hUC-MSCs富含神经营养因子,是一种理想的用于神经损伤治疗的种子细胞.
目的 通過檢測人臍帶間充質榦細胞(hUC-MSCs)的神經營養因子譜,以進一步認識hUC-MSCs通過分泌神經營養因子治療神經損傷的作用. 方法 組織塊培養法分離培養hUC-MSCs,流式細胞儀鑒定榦細胞錶麵標記物,Raybio人細胞因子抗體芯片檢測hUC-MSCs神經營養因子錶達情況,RT-PCR檢測hUC-MSCs神經營養因子的基因錶達,ELISA法定量檢測hUC-MSCs上清液中分泌的腦源性神經營養因子(BDNF)、膠質細胞源性神經營養因子(GDNF)、肝細胞生長因子(HGF)、神經營養因子3(NT-3)含量. 結果 流式細胞分析結果顯示hUC-MSCs錶麵標記物CD29、CD44、CD90暘性(94.05％、90.75％、98.12％),CD34、CD45陰性(3.09％、0.80％).Raybio人細胞因子抗體芯片結果顯示,與神經營養相關的因子BDNF、錶皮生長因子(EGF)、GDNF、HGF、NT-3、血管內皮細胞生長因子(VEGF)、胰島素樣生長因子-1(IGF-1)、神經營養因子4(NT-4)在hUC-MSCs中均有錶達.RT-PCR檢驗也證實IGF-1、VEGF、神經生長因子(NGF)、HGF、GDNF、NT-4、BDNF和NT-3基因在hUC-MSCs錶達.ELISA檢測細胞上清液中BDNF、GDNF、HGF、NT-3的含量分彆為(475.20±32.22) pg/mL、(82.33±3.39) pg/mL、(704.50±12.86) pg/mL、(230.41±16.66) pg/mL,分彆與空白對照組相比,差異均有統計學意義(P＜0.05). 結論 hUC-MSCs富含神經營養因子,是一種理想的用于神經損傷治療的種子細胞.
목적 통과검측인제대간충질간세포(hUC-MSCs)적신경영양인자보,이진일보인식hUC-MSCs통과분비신경영양인자치료신경손상적작용. 방법 조직괴배양법분리배양hUC-MSCs,류식세포의감정간세포표면표기물,Raybio인세포인자항체심편검측hUC-MSCs신경영양인자표체정황,RT-PCR검측hUC-MSCs신경영양인자적기인표체,ELISA법정량검측hUC-MSCs상청액중분비적뇌원성신경영양인자(BDNF)、효질세포원성신경영양인자(GDNF)、간세포생장인자(HGF)、신경영양인자3(NT-3)함량. 결과 류식세포분석결과현시hUC-MSCs표면표기물CD29、CD44、CD90양성(94.05％、90.75％、98.12％),CD34、CD45음성(3.09％、0.80％).Raybio인세포인자항체심편결과현시,여신경영양상관적인자BDNF、표피생장인자(EGF)、GDNF、HGF、NT-3、혈관내피세포생장인자(VEGF)、이도소양생장인자-1(IGF-1)、신경영양인자4(NT-4)재hUC-MSCs중균유표체.RT-PCR검험야증실IGF-1、VEGF、신경생장인자(NGF)、HGF、GDNF、NT-4、BDNF화NT-3기인재hUC-MSCs표체.ELISA검측세포상청액중BDNF、GDNF、HGF、NT-3적함량분별위(475.20±32.22) pg/mL、(82.33±3.39) pg/mL、(704.50±12.86) pg/mL、(230.41±16.66) pg/mL,분별여공백대조조상비,차이균유통계학의의(P＜0.05). 결론 hUC-MSCs부함신경영양인자,시일충이상적용우신경손상치료적충자세포.
Objective To observe the profile of neurotrophic factors in human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) with Raybio antibody chip system,and provide a new cell seed source for spine cord injury or peripheral nerve regeneration. Mcthods The hUC-MSCs were cultured by tissue culture. Characteristics of cell phenotype were confirmed by flow cytometry.Raybio human cytokine antibody array and RT-PCR were employed to detect the neurotropic factors in hUC-MSCs. ELISA was used to determine the contents of growth factors (brain-derived neurotrophic factor [BDNF],glial cell line-derived neurotrophic factor [GDNF],hepatocyte growth factor [HGF],neurotrophin-3 [NT-3]) secreted by hUC-MSCs. Results The hUC-MSCs showed positive results for CD29 (94.05％),CD44 (90.75％) and CD90 (98.12％),and negative results for CD34 (3.09％) and CD45 (0.80％).Array results showed that hUC-MSCs expressed BDNF,epidermal growth factor (EGF),GDNF,HGF and NT-3,vascular endothelial growth factor (VEGF),insulin-like growth factor 1 (IGF-1) and NT-4.The gene expressions ofIGF-1,VEGF,nerve growth factor (NGF),HGF,GDNF,NT-4,BDNF and NT-3 were further verified by RT-PCR.ELISA results showed that the contents of BDNF,GDNF,HGF and NT-3 in the conditioned medium were (475.20±32.22) pg/mL,(82.33±3.39) pg/mL,(704.50±12.86)pg/mL,and (230.41 ±16.66) pg/mL,respectively.These were obviously higher than that in the control group. Conclusion The hUC-MSCs,containing plenty of growth factors,especially neurotrophic factors,may be promising seed cells for spine cord injury and nerve regeneration.
