白来航鸡不同来源间充质干细胞的比较研究

doi:10.16431/j.cnki.1671-7236.2023.10.006

摘要/Abstract

摘要： 【目的】以禽类间充质干细胞(MSCs)为研究对象,探讨不同来源MSCs生物学特性上的差异及其临床应用潜力。【方法】培养白来航鸡4种不同组织器官骨髓(BM)、脂肪(AT)、脐带(UC)和脾脏(S)衍生的MSCs,通过实时荧光定量PCR技术检测细胞表面标记物的表达,采用细胞生长曲线观察种群倍增时间变化,利用噻唑蓝细胞毒性检测法(MTT)确定基质细胞衍生因子1(SDF-1)对不同来源MSCs的细胞毒性,并选用细胞小室检测MSCs体外迁移情况,通过蛋白质印迹法方法确定MSCs体外迁移影响因素,最后选用成骨分化和成脂分化观察不同来源MSCs的体外分化情况。【结果】不同来源的MSCs细胞形态基本一致;细胞表面标记物CD29、CD44、CD71、CD90和CD105等基因呈阳性表达;不同来源的MSCs表面标记物的相对表达量不同,CD29基因的相对表达量基本相同,CD44基因在AT-MSCs中的相对表达量显著高于BM-MSCs(P＜0.05),CD71基因在UC-MSCs中的相对表达量显著低于BM-MSCs(P＜0.05),CD90基因在UC-MSCs和S-MSCs中的相对表达量极显著低于BM-MSCs(P＜0.01);CD105基因在S-MSCs中的相对表达量显著低于BM-MSCs(P＜0.05)。4种来源MSCs均有较好的体外生长潜力,其中S-MSCs生产潜伏期最长(40 h)。4种来源MSCs均具有较强的体外迁移能力,S-MSCs中CXC趋化因子受体4(CXCR4)的低表达可能是导致其体外迁移效率较低的原因。4种来源的MSCs均具有体外成骨分化和成脂分化能力,其中BM-MSCs的成骨分化能力较强,S-MSCs的成脂分化能力较强。【结论】白来航鸡4种来源的组织器官均可衍生出MSCs,且均具有良好的体外增殖、体外迁移及体外分化能力。4种MSCs的生物学特性存在差异,具有不同的临床应用潜能,BM-MSCs的综合性能较优,S-MSCs的脂肪诱导分化方面优势明显。本研究加强了对禽类MSCs的了解,对MSCs治疗禽类相关疾病的研究及珍稀禽类的保护具有重要意义。

关键词: 白来航鸡; 间充质干细胞; 表面标记物; 细胞分化; 细胞迁移

Abstract: 【Objective】 Avian mesenchymal stem cells (MSCs) were taken as the research target to investigate the differences in biological characteristics and clinical application potential of MSCs from different sources.【Method】 MSCs derived from bone marrow(BM),adipose tissue(AT),umbilical cord (UC) and spleen (S) of four different tissues and organs in White Leghorn chickens were cultured.The expression of cell surface markers was detected by Real-time quantitative PCR,and the population doubling time was observed by cell growth curve.The cytotoxicity of SDF-1 to MSCs from different sources were determined by thiazole blue cytotoxicity assay (MTT),and the in vitro migration of MSCs was detected by Transwell test.The influencing factors of MSCs migration in vitro were determined by Western blotting.Finally,in vitro differentiation of MSCs from different sources were observed by osteogenic differentiation and lipogenic differentiation.【Result】 The morphology of MSCs from different sources were basically consistent.The marker CD29,CD44,CD71,CD90 and CD105 genes were positively expressed.The relative expression of surface markers in MSCs from different sources were different,and the relative expression of CD29 gene was basically the same,the relative expression of CD44 gene in AT-MSCs was significantly higher than that in BM-MSCs (P＜0.05),and the relative expression of CD71 gene in UC-MSCs was significantly lower than that in BM-MSCs (P＜0.05).The relative expression of CD90 gene in UC-MSCs and S-MSCs was significantly lower than that in BM-MSCs (P＜0.01).The relative expression of CD105 gene in S-MSCs was significantly lower than that in BM-MSCs (P＜0.05).The four sources of MSCs all had good growth potential in vitro,with the longest incubation period of 40 h for S-MSCs.All the MSCs from the four sources had strong in vitro migration ability,and the low expression of CXC-chemokine receptor 4(CXCR4) in S-MSCs might be the cause of their low in vitro migration efficiency.MSCs from the four sources all showed in vitro osteogenic and adipogenic differentiation abilities,among which BM-MSCs had strong osteogenic differentiation ability,while S-MSCs had strong adipogenic differentiation ability.【Conclusion】 MSCs could be derived from four sources of tissues and organs in White Lehang chickens,and all had good in vitro proliferation,migration and differentiation ability.The biological characteristics of the four MSCs were different,inducing different clinical application potential.BM-MSCs had superior comprehensive performance,while S-MSCs had significant advantages in adipose induction differentiation.This study strengthened the understanding of avian MSCs,which was of great significance for the study of MSCs treatment of avian related diseases and the protection of rare birds.

Key words: White Leghorn chickens; mesenchymal stem cells; surface markers; cell differentiation; cell migration

中图分类号:

S831

秦欣欣, 滕文熙, 张翔, 杨雅淋, 胡玉娜, 王东羽, 芮磊, 李赞东. 白来航鸡不同来源间充质干细胞的比较研究[J]. 中国畜牧兽医, 2023, 50(10): 3928-3938.

QIN Xinxin, TENG Wenxi, ZHANG Xiang, YANG Yalin, HU Yuna, WANG Dongyu, RUI Lei, LI Zandong. Comparative Study on Different Sources of Mesenchymal Stem Cells from White Leghorn Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 3928-3938.

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