Expansion of Human Limbal Epithelial Stem/Progenitor Cells Using Different Human Sera: A Multivariate Statistical Analysis.
Expansion of Human Limbal Epithelial Stem/Progenitor Cells Using Different Human Sera: A Multivariate Statistical Analysis.
复制标题
使用不同的人类血清扩展人类边缘上皮干/祖细胞：多元统计分析。
DOI：
10.3390/ijms21176132
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发表时间：
2020-08-25
影响因子：
5.6
通讯作者：
Andollo N
中科院分区：
文献类型：
--
作者：
Hernáez-Moya R;González S;Urkaregi A;Pijoan JI;Deng SX;Andollo N
人类培养的角膜缘上皮干细胞/祖细胞（LESCs）移植已被证明可以恢复约76%角膜缘干细胞缺乏症患者角膜表面的完整性和功能。然而，扩大LESCs有不同的方案，其中许多使用异种产品，对患者的健康构成风险。我们比较了角膜缘外植体在羊膜上的培养，在补充激素上皮培养基（SHEM）中，补充FBS或两种不同产生的人血清。检测细胞形态、细胞大小、细胞生长速度、分化表达水平和推测的干细胞标志物。在人血清中定量测定了几种生物活性分子。在一种新颖的方法中，我们对数据进行了多变量统计分析，以研究培养因素，如特异性影响细胞表型的人类血清中不同表达的分子。我们的研究结果表明，与FBS培养相比，人血清培养的角膜缘细胞生长速度更快，含有相似数量的小细胞，p63α蛋白的表达更高，细胞角蛋白K12的表达更低，从而保持了LESCs的干/祖表型。此外，多变量分析提供了大量数据，以更好地理解由于使用不同的培养方法或不同的培养成分而获得不同的细胞表型。
Transplantation of human cultured limbal epithelial stem/progenitor cells (LESCs) has demonstrated to restore the integrity and functionality of the corneal surface in about 76% of patients with limbal stem cell deficiency. However, there are different protocols for the expansion of LESCs, and many of them use xenogeneic products, being a risk for the patients’ health. We compared the culture of limbal explants on the denuded amniotic membrane in the culture medium—supplemental hormone epithelial medium (SHEM)—supplemented with FBS or two differently produced human sera. Cell morphology, cell size, cell growth rate, and the expression level of differentiation and putative stem cell markers were examined. Several bioactive molecules were quantified in the human sera. In a novel approach, we performed a multivariate statistical analysis of data to investigate the culture factors, such as differently expressed molecules of human sera that specifically influence the cell phenotype. Our results showed that limbal cells cultured with human sera grew faster and contained similar amounts of small-sized cells, higher expression of the protein p63α, and lower of cytokeratin K12 than FBS cultures, thus, maintaining the stem/progenitor phenotype of LESCs. Furthermore, the multivariate analysis provided much data to better understand the obtaining of different cell phenotypes as a consequence of the use of different culture methodologies or different culture components.
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DOI：
10.1089/ten.tec.2016.0388
发表时间：
2017-04
期刊：
Tissue engineering. Part C, Methods
影响因子：
--
作者：
González S;Chen L;Deng SX
通讯作者：
Deng SX
影响因子：
3.4
作者：
GARCIAHIRSCHFELD, J;LOPEZBRIONES, LG;BELMONTE, C
通讯作者：
BELMONTE, C
影响因子：
4.4
作者：
Freire, Vanesa;Andollo, Noelia;Morales, Maria-Celia
通讯作者：
Morales, Maria-Celia
影响因子：
2.8
作者：
Behaegel, Josephine;Zakaria, Nadia;Dhubhghaill, Sorcha Ni
通讯作者：
Dhubhghaill, Sorcha Ni
影响因子：
1.2
作者：
Gonzalez, Sheyla;Mei, Hua;Deng, Sophie X.
通讯作者：
Deng, Sophie X.