Abstract

Background: Human platelet lysates (HPLs) are emerging as the new gold standard supplement of growth media for ex vivo expansion of cells for transplant. However, variations do exist in the way how HPLs are prepared. In particular, uncertainties still exist regarding the type of HPL most suitable for corneal endothelium cells (CEC) expansion, especially as these cells have limited proliferative capacity. Material and methods: Three distinct HPL preparations were produced, with or without calcium chloride/glass beads activation, and with or without heat treatment at 56. °C for 30. min. These HPLs were used to supplement basal D-MEM growth medium, each at a protein concentration equivalent to that of 10% fetal bovine serum (FBS; control). Impact on CEC (BCE C/D-1b cells) in vitro morphology, viability and capacity to express Zonula occludens-1 (ZO-1) tight junction marker was assessed by Western blotting. Results: BCE C/D-1b cells grown in all HPL supplements exhibited four of essential characteristic properties: adhesion capacity, microscopic morphology and viability similar to that observed when using 10% FBS. In addition, Western blots analysis revealed an expression of the ZO-1 marker by BCE C/D-1b cells in all conditions of culture. Conclusion: CECs can expand ex vivo in a basal medium supplemented with the three HPLs without noticeable difference compared to FBS supplement. These data support further studies to evaluate the potential to use HPLs as a clinical-grade xeno-free supplement of CEC for corneal transplant.

Original languageEnglish
JournalTransfusion and Apheresis Science
DOIs
Publication statusPublished - 2017

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Corneal Endothelium
Blood Platelets
Tight Junctions
Western Blotting
Transplants
Calcium Chloride
In Vitro Techniques
Growth
Uncertainty
Glass
Hot Temperature

Keywords

  • Corneal endothelium
  • Expansion
  • Fetal bovine serum
  • Platelet lysate

ASJC Scopus subject areas

  • Hematology

Cite this

@article{0e66ab54d8424cb093084cf1cfd151e4,
title = "Comparison of three human platelet lysates used as supplements for in vitro expansion of corneal endothelium cells",
abstract = "Background: Human platelet lysates (HPLs) are emerging as the new gold standard supplement of growth media for ex vivo expansion of cells for transplant. However, variations do exist in the way how HPLs are prepared. In particular, uncertainties still exist regarding the type of HPL most suitable for corneal endothelium cells (CEC) expansion, especially as these cells have limited proliferative capacity. Material and methods: Three distinct HPL preparations were produced, with or without calcium chloride/glass beads activation, and with or without heat treatment at 56. °C for 30. min. These HPLs were used to supplement basal D-MEM growth medium, each at a protein concentration equivalent to that of 10{\%} fetal bovine serum (FBS; control). Impact on CEC (BCE C/D-1b cells) in vitro morphology, viability and capacity to express Zonula occludens-1 (ZO-1) tight junction marker was assessed by Western blotting. Results: BCE C/D-1b cells grown in all HPL supplements exhibited four of essential characteristic properties: adhesion capacity, microscopic morphology and viability similar to that observed when using 10{\%} FBS. In addition, Western blots analysis revealed an expression of the ZO-1 marker by BCE C/D-1b cells in all conditions of culture. Conclusion: CECs can expand ex vivo in a basal medium supplemented with the three HPLs without noticeable difference compared to FBS supplement. These data support further studies to evaluate the potential to use HPLs as a clinical-grade xeno-free supplement of CEC for corneal transplant.",
keywords = "Corneal endothelium, Expansion, Fetal bovine serum, Platelet lysate",
author = "Wang, {Tsung Jen} and Chen, {Ming Sheng} and Chou, {Ming Li} and Lin, {Hsiu Chen} and Jerard Seghatchian and Thierry Burnouf",
year = "2017",
doi = "10.1016/j.transci.2017.08.021",
language = "English",
journal = "Transfusion and Apheresis Science",
issn = "1473-0502",
publisher = "Elsevier Limited",

}

TY - JOUR

T1 - Comparison of three human platelet lysates used as supplements for in vitro expansion of corneal endothelium cells

AU - Wang, Tsung Jen

AU - Chen, Ming Sheng

AU - Chou, Ming Li

AU - Lin, Hsiu Chen

AU - Seghatchian, Jerard

AU - Burnouf, Thierry

PY - 2017

Y1 - 2017

N2 - Background: Human platelet lysates (HPLs) are emerging as the new gold standard supplement of growth media for ex vivo expansion of cells for transplant. However, variations do exist in the way how HPLs are prepared. In particular, uncertainties still exist regarding the type of HPL most suitable for corneal endothelium cells (CEC) expansion, especially as these cells have limited proliferative capacity. Material and methods: Three distinct HPL preparations were produced, with or without calcium chloride/glass beads activation, and with or without heat treatment at 56. °C for 30. min. These HPLs were used to supplement basal D-MEM growth medium, each at a protein concentration equivalent to that of 10% fetal bovine serum (FBS; control). Impact on CEC (BCE C/D-1b cells) in vitro morphology, viability and capacity to express Zonula occludens-1 (ZO-1) tight junction marker was assessed by Western blotting. Results: BCE C/D-1b cells grown in all HPL supplements exhibited four of essential characteristic properties: adhesion capacity, microscopic morphology and viability similar to that observed when using 10% FBS. In addition, Western blots analysis revealed an expression of the ZO-1 marker by BCE C/D-1b cells in all conditions of culture. Conclusion: CECs can expand ex vivo in a basal medium supplemented with the three HPLs without noticeable difference compared to FBS supplement. These data support further studies to evaluate the potential to use HPLs as a clinical-grade xeno-free supplement of CEC for corneal transplant.

AB - Background: Human platelet lysates (HPLs) are emerging as the new gold standard supplement of growth media for ex vivo expansion of cells for transplant. However, variations do exist in the way how HPLs are prepared. In particular, uncertainties still exist regarding the type of HPL most suitable for corneal endothelium cells (CEC) expansion, especially as these cells have limited proliferative capacity. Material and methods: Three distinct HPL preparations were produced, with or without calcium chloride/glass beads activation, and with or without heat treatment at 56. °C for 30. min. These HPLs were used to supplement basal D-MEM growth medium, each at a protein concentration equivalent to that of 10% fetal bovine serum (FBS; control). Impact on CEC (BCE C/D-1b cells) in vitro morphology, viability and capacity to express Zonula occludens-1 (ZO-1) tight junction marker was assessed by Western blotting. Results: BCE C/D-1b cells grown in all HPL supplements exhibited four of essential characteristic properties: adhesion capacity, microscopic morphology and viability similar to that observed when using 10% FBS. In addition, Western blots analysis revealed an expression of the ZO-1 marker by BCE C/D-1b cells in all conditions of culture. Conclusion: CECs can expand ex vivo in a basal medium supplemented with the three HPLs without noticeable difference compared to FBS supplement. These data support further studies to evaluate the potential to use HPLs as a clinical-grade xeno-free supplement of CEC for corneal transplant.

KW - Corneal endothelium

KW - Expansion

KW - Fetal bovine serum

KW - Platelet lysate

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