Expansion of Human Limbal Epithelial Stem/Progenitor Cells Using Different Human Sera: A Multivariate Statistical Analysis

被引:7
作者
Hernaez-Moya, Raquel [1 ]
Gonzalez, Sheyla [2 ]
Urkaregi, Arantza [3 ]
Ignacio Pijoan, Jose [4 ]
Deng, Sophie X. [2 ]
Andollo, Noelia [1 ]
机构
[1] Univ Basque Country UPV EHU, Sch Med & Nursing, Biocruces Bizkaia Hlth Res Inst, Dept Cell Biol & Histol, Leioa 48940, Bizkaia, Spain
[2] Univ Calif Los Angeles, Stein Eye Inst, Cornea Div, Los Angeles, CA 90095 USA
[3] Univ Basque Country UPV EHU, Biocruces Bizkaia Hlth Res Inst, Dept Appl Math & Stat & Operat Res, Leioa 48940, Bizkaia, Spain
[4] Cruces Univ Hosp, Biocruces Bizkaia Hlth Res Inst, Clin Epidemiol Unit, Baracaldo 48903, Bizkaia, Spain
关键词
limbal epithelial stem; progenitor cells (LESC); limbal stem cell deficiency (LSCD); cell therapy; xenogeneic-free cell expansion; human serum (HS); serum derived from plasma rich in growth factors (s-PRGF); human amniotic membrane (HAM); explants culture; multivariate statistical analysis; principal component analysis (PCA) combined with clusters analysis; partial least squares discriminant analysis (PLS-DA); EX-VIVO EXPANSION; 3 BLOOD DERIVATIVES; NERVE GROWTH-FACTOR; FREE CULTURE-SYSTEM; FETAL BOVINE SERUM; SUBSTANCE-P; STEM-CELLS; AMNIOTIC MEMBRANE; STROMAL CELLS; IN-VITRO;
D O I
10.3390/ijms21176132
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 p63 alpha, 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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页码:1 / 24
页数:24
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