The North West Embryonic Stem Cell Centre

Establishment of GMP hES cell lines

Our work in this area aims to establish standardised methods for maintenance of pluripotency using existing human Embryonic Stem (hES) cell lines (HES-3, HUES-3 and -7) and applying these to the derivation of new hESC lines.

Researchers at NWESCC have established a standardised protocol for enzymatic passaging on existing hES lines. They are also making progress on xenofree passaging. Both techniques are essential for GMP derivation of clinical grade hES cells. Embryos are donated by IVF patients at St Mary’s Hospital with consent for hES cell derivation. This has culminated in the successful derivation of our first human ES cell line, designated MAN-1, which we have now characterised. This supply of embryos has also allowed us to refine inner cell mass isolation from the blastocyst in the absence of xenobiotics.

In order to eliminate the use of non-human contaminants, we have developed a collaboration with Prof John Aplin at St. Mary’s Hospital Medical School, Manchester, to develop human placental stromal fibroblast (PSF) feeder cells. We will use these cells to grow our established and then newly derived hES cell lines. We have initiated the process of defining a GMP protocol for screening, passaging and stockpiling PSF cells for deriving and culturing future MAN hES cell lines. Our goal is to utilise such a protocol in order to generate a GMP immortalised PSF line using integrating viral vectors. This line, when established, will allow us to attain reproducibility between ongoing derivations and during culture.

Principal investigators

Postdoctoral research associate

Quality management

  • Miss Alexandra Ross

Research technicians

Establish protocols

Human blastocyst
Researchers at NWESCC have established a standardised protocol for enzymatic passaging on existing hES lines.