Gene expression profiles of the meniscus avascular phenotype: A guide for meniscus tissue engineering.
Gene expression profiles of the meniscus avascular phenotype: A guide for meniscus tissue engineering.
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DOI：
10.1002/jor.23864
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发表时间：
2018-07
期刊：
影响因子：
--
通讯作者：
D'Lima DD
中科院分区：
文献类型：
--
作者：
Grogan SP;Duffy SF;Pauli C;Lotz MK;D'Lima DD
无血管（Avas）半月板再生仍然是一个挑战，这在一定程度上是我们对维持该组织区域的细胞知识有限的结果。在这项研究中，我们利用微阵列来表征完整的人类Avas半月板组织和培养扩增后的细胞的基因表达谱。使用这些数据，我们检查了各种3D培养条件，以使Avas细胞向组织表型再分化。直接从组织中分离RNA，或在细胞分离和单层培养2周后分离RNA。RNA在人类基因组阵列上杂交。通过排序分析鉴定差异表达基因。进行大卫途径分析并使用STRING分析可视化。使用对另外的供体组织（组织和细胞）的定量PCR（qPCR）来验证阵列数据。对3D培养的Avas细胞进行qPCR，以与阵列生成的数据进行比较。基于分化状态，总共387个基因是DE（>3倍变化; p < 0.01）。在Avas培养的细胞中，上调的途径包括粘着斑、ECM-受体相互作用、肌动蛋白细胞骨架的调节和PDGF信号传导。在三维培养的Avas细胞中，TGFβ1或TGFβ1和BMP 6的组合对促进Avas组织表型最有效。THBS 2和THBS 4表达水平被鉴定为表示半月板细胞表型状态的手段。我们确定了关键的基因表达谱，新的标志物和途径参与表征的Avas半月板表型在自然状态下，在体外脱分化和再分化。这些数据用于筛选3D条件以产生类牙龈瘤样新组织。
Avascular (Avas) meniscus regeneration remains a challenge, which is partly a consequence of our limited knowledge of the cells that maintain this tissue region. In this study, we utilized microarrays to characterize gene expression profiles of intact human Avas meniscus tissue and of cells following culture expansion. Using these data, we examined various 3D culture conditions to redifferentiate Avas cells toward the tissue phenotype. RNA was isolated from either the tissue directly or following cell isolation and 2 weeks in monolayer culture. RNA was hybridized on human genome arrays. Differentially expressed (DE) genes were identified by ranking analysis. DAVID pathway analysis was performed and visualized using STRING analysis. Quantitative PCR (qPCR) on additional donor menisci (tissues and cells) were used to validate array data. Avas cells cultured in 3D were subjected to qPCR to compare with the array-generated data. A total of 387 genes were DE based on differentiation state (>3-fold change; p < 0.01). In Avas cultured cells, the upregulated pathways included focal adhesion, ECM-receptor interaction, regulation of actin cytoskeleton, and PDGF Signaling. In 3D-cultured Avas cells, TGFβ1 or combinations of TGFβ1 and BMP6 were most effective to promote an Avas tissue phenotype. THBS2 and THBS4 expression levels were identified as a means to denote meniscus cell phenotype status. We identified the key gene expression profiles, new markers and pathways involved in characterizing the Avas meniscus phenotype in the native state and during in vitro dedifferentiation and redifferentiation. These data served to screen 3D conditions to generate meniscus-like neotissues.
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影响因子：
4.1
作者：
Baker, Brendon M.;Shah, Roshan P.;Mauck, Robert L.
通讯作者：
Mauck, Robert L.
DOI：
10.1083/jcb.138.5.1159
发表时间：
1997-09-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Camper L;Heinegârd D;Lundgren-Akerlund E
通讯作者：
Lundgren-Akerlund E
影响因子：
6
作者：
Biswas, Saptarshi;Bachay, Galina;Brunken, William J.
通讯作者：
Brunken, William J.
DOI：
10.1016/j.arthro.2011.06.041
发表时间：
2011-12-01
影响因子：
4.7
作者：
Angel Ruiz-Iban, Miguel;Diaz-Heredia, Jorge;Abraira, Victor
通讯作者：
Abraira, Victor
影响因子：
64.5
作者：
BENYA, PD;PADILLA, SR;NIMNI, ME
通讯作者：
NIMNI, ME