Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome

Jui-Fang Tsai, Ryan Yao Nien Tsai

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Objectives: To identify the stem-cell property of the ex vivo expansion of limbal stem cells (LSCs) on amniotic membrane (AM) in culture system and after clinical transplantation. Methods: Four key factors have to be performed in the defined culture system: (1) the label-retaining cells have to be identified; (2) the cells can be serially expanded and passaged in vitro; (3) the expanded cells can be labeled by tissue-specific keratin or markers, and (3) their stem cells cannot be labeled by those keratin or markers. Results: The ex vivo-expanded LSCs on AM were positive for p63 and ABCG2 and BrdU label-retaining studies on flat mount preparation. When the ex vivo-expanded LSCs with AM were transplanted into a subcutaneous layer of nude mice, they formed multiple layers of cells. Only the basal layer of cells was positive for p63 and BrdU. The cells over the suprabasal layers were positive for K12/K3. The pathologic studies of corneal specimen of successful LSC transplantation after penetrating keratoplasty demonstrated that P63-positive cells were noticed all over the basal layer of central cornea and AM could be identified at 10 months after LSC transplantation. Conclusions: These results indicate that the AM provided the niche function for cultured LSCs and maintained the limbal-like environment for the transplanted area of cornea. The survival of cases depends on the severity of the disease entity, culture technique, and maintenance of the niche environment for LSCs in the culture and after clinical transplantation.

Original languageEnglish
Pages (from-to)305-309
Number of pages5
JournalEye and Contact Lense
Volume36
Issue number5
DOIs
Publication statusPublished - Sep 2010

Fingerprint

Amnion
Stem Cells
Stem Cell Transplantation
Bromodeoxyuridine
Keratins
Cornea
Transplantation
Culture Techniques
Penetrating Keratoplasty
Nude Mice
Cell Culture Techniques
Maintenance

Keywords

  • Amniotic membrane
  • Ex vivo expansion
  • Limbal stem cells
  • Niche

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome. / Tsai, Jui-Fang; Tsai, Ryan Yao Nien.

In: Eye and Contact Lense, Vol. 36, No. 5, 09.2010, p. 305-309.

Research output: Contribution to journalArticle

Tsai, Jui-Fang ; Tsai, Ryan Yao Nien. / Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome. In: Eye and Contact Lense. 2010 ; Vol. 36, No. 5. pp. 305-309.
@article{a708cce5ac1645148fec5a594724dc27,
title = "Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome",
abstract = "Objectives: To identify the stem-cell property of the ex vivo expansion of limbal stem cells (LSCs) on amniotic membrane (AM) in culture system and after clinical transplantation. Methods: Four key factors have to be performed in the defined culture system: (1) the label-retaining cells have to be identified; (2) the cells can be serially expanded and passaged in vitro; (3) the expanded cells can be labeled by tissue-specific keratin or markers, and (3) their stem cells cannot be labeled by those keratin or markers. Results: The ex vivo-expanded LSCs on AM were positive for p63 and ABCG2 and BrdU label-retaining studies on flat mount preparation. When the ex vivo-expanded LSCs with AM were transplanted into a subcutaneous layer of nude mice, they formed multiple layers of cells. Only the basal layer of cells was positive for p63 and BrdU. The cells over the suprabasal layers were positive for K12/K3. The pathologic studies of corneal specimen of successful LSC transplantation after penetrating keratoplasty demonstrated that P63-positive cells were noticed all over the basal layer of central cornea and AM could be identified at 10 months after LSC transplantation. Conclusions: These results indicate that the AM provided the niche function for cultured LSCs and maintained the limbal-like environment for the transplanted area of cornea. The survival of cases depends on the severity of the disease entity, culture technique, and maintenance of the niche environment for LSCs in the culture and after clinical transplantation.",
keywords = "Amniotic membrane, Ex vivo expansion, Limbal stem cells, Niche",
author = "Jui-Fang Tsai and Tsai, {Ryan Yao Nien}",
year = "2010",
month = "9",
doi = "10.1097/ICL.0b013e3181efff40",
language = "English",
volume = "36",
pages = "305--309",
journal = "Eye and Contact Lens",
issn = "1542-2321",
publisher = "Lippincott Williams and Wilkins",
number = "5",

}

TY - JOUR

T1 - Ex vivo expansion of corneal stem cells on amniotic membrane and their outcome

AU - Tsai, Jui-Fang

AU - Tsai, Ryan Yao Nien

PY - 2010/9

Y1 - 2010/9

N2 - Objectives: To identify the stem-cell property of the ex vivo expansion of limbal stem cells (LSCs) on amniotic membrane (AM) in culture system and after clinical transplantation. Methods: Four key factors have to be performed in the defined culture system: (1) the label-retaining cells have to be identified; (2) the cells can be serially expanded and passaged in vitro; (3) the expanded cells can be labeled by tissue-specific keratin or markers, and (3) their stem cells cannot be labeled by those keratin or markers. Results: The ex vivo-expanded LSCs on AM were positive for p63 and ABCG2 and BrdU label-retaining studies on flat mount preparation. When the ex vivo-expanded LSCs with AM were transplanted into a subcutaneous layer of nude mice, they formed multiple layers of cells. Only the basal layer of cells was positive for p63 and BrdU. The cells over the suprabasal layers were positive for K12/K3. The pathologic studies of corneal specimen of successful LSC transplantation after penetrating keratoplasty demonstrated that P63-positive cells were noticed all over the basal layer of central cornea and AM could be identified at 10 months after LSC transplantation. Conclusions: These results indicate that the AM provided the niche function for cultured LSCs and maintained the limbal-like environment for the transplanted area of cornea. The survival of cases depends on the severity of the disease entity, culture technique, and maintenance of the niche environment for LSCs in the culture and after clinical transplantation.

AB - Objectives: To identify the stem-cell property of the ex vivo expansion of limbal stem cells (LSCs) on amniotic membrane (AM) in culture system and after clinical transplantation. Methods: Four key factors have to be performed in the defined culture system: (1) the label-retaining cells have to be identified; (2) the cells can be serially expanded and passaged in vitro; (3) the expanded cells can be labeled by tissue-specific keratin or markers, and (3) their stem cells cannot be labeled by those keratin or markers. Results: The ex vivo-expanded LSCs on AM were positive for p63 and ABCG2 and BrdU label-retaining studies on flat mount preparation. When the ex vivo-expanded LSCs with AM were transplanted into a subcutaneous layer of nude mice, they formed multiple layers of cells. Only the basal layer of cells was positive for p63 and BrdU. The cells over the suprabasal layers were positive for K12/K3. The pathologic studies of corneal specimen of successful LSC transplantation after penetrating keratoplasty demonstrated that P63-positive cells were noticed all over the basal layer of central cornea and AM could be identified at 10 months after LSC transplantation. Conclusions: These results indicate that the AM provided the niche function for cultured LSCs and maintained the limbal-like environment for the transplanted area of cornea. The survival of cases depends on the severity of the disease entity, culture technique, and maintenance of the niche environment for LSCs in the culture and after clinical transplantation.

KW - Amniotic membrane

KW - Ex vivo expansion

KW - Limbal stem cells

KW - Niche

UR - http://www.scopus.com/inward/record.url?scp=77957574238&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957574238&partnerID=8YFLogxK

U2 - 10.1097/ICL.0b013e3181efff40

DO - 10.1097/ICL.0b013e3181efff40

M3 - Article

C2 - 20823708

AN - SCOPUS:77957574238

VL - 36

SP - 305

EP - 309

JO - Eye and Contact Lens

JF - Eye and Contact Lens

SN - 1542-2321

IS - 5

ER -