Open AccessDerivation of Xeno-Free and GMP-Grade Human Embryonic Stem Cells – Platforms for Future Clinical Applications
Author(s) -
Shelly E. Tannenbaum,
Tikva Turetsky,
Orna Singer,
Einat Aizenman,
Sophie Kirshberg,
Nili Ilouz,
Yaniv Gil,
Yael Berman-Zaken,
Temima Schnitzer Perlman,
Nitshia Geva,
Ora Levy,
Daniel Arbell,
Alex Simon,
Assaf BenMeir,
Yoel Shufaro,
Neri Laufer,
Benjamin Reubinoff
Publication year - 2012
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0035325
Subject(s) - embryonic stem cell , regenerative medicine , stem cell , microbiology and biotechnology , umbilical cord , induced pluripotent stem cell , biology , good manufacturing practice , computational biology , biomedical engineering , immunology , medicine , genetics , regulatory affairs , gene
Clinically compliant human embryonic stem cells (hESCs) should be developed in adherence to ethical standards, without risk of contamination by adventitious agents. Here we developed for the first time animal-component free and good manufacturing practice (GMP)-compliant hESCs. After vendor and raw material qualification, we derived xeno-free, GMP-grade feeders from umbilical cord tissue, and utilized them within a novel, xeno-free hESC culture system. We derived and characterized three hESC lines in adherence to regulations for embryo procurement, and good tissue, manufacturing and laboratory practices. To minimize freezing and thawing, we continuously expanded the lines from initial outgrowths and samples were cryopreserved as early stocks and banks. Batch release criteria included DNA-fingerprinting and HLA-typing for identity, characterization of pluripotency-associated marker expression, proliferation, karyotyping and differentiation in-vitro and in-vivo. These hESCs may be valuable for regenerative therapy. The ethical, scientific and regulatory methodology presented here may serve for development of additional clinical-grade hESCs.