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Substitutes of fetal bovine sera for the culture of human corneal limbal stem cells
Author(s) -
IÑIGO PORTUGUÉS A,
MEANA A,
VÁZQUEZ N,
CHACÓN M,
RIESTRA ANAC,
ORIVE G,
ANITUA E,
MERAYOLLOVES J
Publication year - 2014
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2014.s053.x
Subject(s) - fetal bovine serum , cytokeratin , explant culture , stem cell , immunocytochemistry , andrology , cell culture , biology , cell growth , fetus , immunology , cell , pathology , microbiology and biotechnology , in vitro , medicine , biochemistry , immunohistochemistry , pregnancy , genetics
Purpose We analyze the potential use of PRGF‐Endoret® and human sera (HS) as a substitute of xenogeneic sera in corneal cell cultures. Methods Plasma rich in growth factors (PRGF) eye drop and HS was obtained at our local blood bank from human blood of healthy donors after informed consent. Briefly PRGF was purified using the Endoret kit in Ophthalmology. HS was obtained according to standardized protocols. Limbal stem cells were obtained from explants of 2‐3mm in diameter of the limbal region of corneoscleral tissue and cultured in culture medium containing 10% PRGF‐Endoret®, HS or fetal bovine serum (FBS) for 7 days. Afterwards, cells were fixed and growth area was quantified. Immunocytochemistry for p.63, ABCG‐2 and cytokeratin was also performed in order to check their immunological markers. Results PRGF‐Endoret®, HS and FBS, cell cultures showed a positive expression in p.63, ABCG‐2 and cytokeratin. However some differences were found between cultures, limbal stem cell cultures supplemented with PRGF‐Endoret® and HS showed a better growth area in contrast with the ones supplemented with FBS. Moreover, there was cellular growth in more of the explants cultured in PRGF‐Endoret® and HS compared to FBS supplemented cultures. Conclusion PRGF‐Endoret® and HS seems to improve the cellular growth and, while maintaining the p63 and ABCG‐2 expression compared to FBS supplemented cultures. Therefore, PRGF‐Endoret® or HS could be used as a substitute of xenogeneic sera, not only as an allogeneic product but also as an autologous product for the culture and expansion of corneal limbal stem cells.