Ascorbate-dependent impact on cell-derived matrix in modulation of stiffness and rejuvenation of infrapatellar fat derived stem cells toward chondrogenesis.
Ascorbate-dependent impact on cell-derived matrix in modulation of stiffness and rejuvenation of infrapatellar fat derived stem cells toward chondrogenesis.
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DOI：
10.1088/1748-6041/11/4/045009
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发表时间：
2016-08-10
期刊：
影响因子：
--
通讯作者：
Pei M
中科院分区：
文献类型：
--
作者：
Pizzute T;Zhang Y;He F;Pei M
利用细胞衍生的脱细胞细胞外基质（dECM）开发体外微环境是有效扩增用于软骨工程和再生的成体干细胞的有前途的方法。抗坏血酸作为细胞合成胶原蛋白的关键刺激物，胶原蛋白构成dECM的主要成分。在这项研究中，我们假设，优化抗坏血酸治疗将最大限度地提高再生效果的dECM对扩增的干细胞从人髌下脂肪垫在增殖和软骨分化。在持续时间方案的研究中，我们发现，dECM没有L-抗坏血酸磷酸盐（AA）的治疗，表现出较低的刚度测量原子力显微镜，产生了更高的增殖能力，但较低的软骨形成的潜力相比，那些具有不同的AA治疗持续时间的扩展细胞。如果沉积的细胞来自第2代而不是第5代，则用250 μM AA处理10天的dECM在软骨形成能力方面具有更好的复壮，尽管基质硬度没有显著差异。在剂量方案研究中，我们发现，由不同浓度的AA沉积的dECM产生具有较高增殖能力的扩增细胞，尽管干细胞相关表面标志物的表达水平较低。与在组织培养聚苯乙烯上扩增的细胞相比，在dECM上的细胞表现出更大的软骨形成潜力，特别是对于50 μM和250 μM AA处理的dECM。在补充3，4-二羟基苯甲酸乙酯（EDHB）（一种靶向前胶原合成的抑制剂）的情况下，50 μM AA处理的dECM在mRNA和蛋白质水平上显示出扩增细胞软骨形成潜力的复壮效应显著降低，尽管基质硬度没有显著差异。在基质制备过程中定义的AA治疗将有利于dECM介导的干细胞工程和未来的软骨缺损治疗。
Developing an in vitro microenvironment using cell-derived decellularized extracellular matrix (dECM) is a promising approach to efficiently expand adult stem cells for cartilage engineering and regeneration. Ascorbic acid serves as a critical stimulus for cells to synthesize collagens, which constitute the major component of dECM. In this study, we hypothesized that optimization of ascorbate treatment would maximize the rejuvenation effect of dECM on expanded stem cells from human infrapatellar fat pad in both proliferation and chondrogenic differentiation. In the duration regimen study, we found that dECM without L-ascorbic acid phosphate (AA) treatment, exhibiting lower stiffness measured by atomic force microscopy, yielded expanded cells with higher proliferation capacity but lower chondrogenic potential when compared to those with varied durations of AA treatment. dECM with 250 μM of AA treatment for 10 days had better rejuvenation in chondrogenic capacity if the deposited cells were from passage 2 rather than passage 5, despite no significant difference in matrix stiffness. In the dose regimen study, we found that dECMs deposited by varied concentrations of AA yielded expanded cells with higher proliferation capacity despite lower expression levels of stem cell related surface markers. Compared to cells expanded on tissue culture polystyrene, those on dECM exhibited greater chondrogenic potential, particularly for the dECMs with 50 μM and 250 μM of AA treatment. With the supplementation of ethyl-3,4-dihydroxybenzoate (EDHB), an inhibitor targeting procollagen synthesis, the dECM with 50 μM of AA treatment exhibited a dramatic decrease in the rejuvenation effect of expanded cell chondrogenic potential at both mRNA and protein levels despite no significant difference in matrix stiffness. Defined AA treatments during matrix preparation will benefit dECM-mediated stem cell engineering and future treatments for cartilage defects.
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影响因子：
14
作者：
Li, Jingting;Hansen, Kirk C.;Pei, Ming
通讯作者：
Pei, Ming
DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
影响因子：
3.1
作者：
Altaf, F. M.;Hering, T. M.;Johnstone, B.
通讯作者：
Johnstone, B.
影响因子：
3.8
作者：
Bhana, Bashir;Iyer, Rohin K.;Radisic, Milica
通讯作者：
Radisic, Milica
影响因子：
--
作者：
Mochizuki, T;Muneta, T;Sekiya, I
通讯作者：
Sekiya, I