Laminin 521 stabilizes the pluripotency expression pattern of human embryonic stem cells initially derived on feeder cells

In this article, the authors tested the effect of the laminin-521 substrate on cultured hES cells. Five male hES cell lines, originally derived on human foreskin fibroblasts (hFFs), were cultured on hFF, Matrigel, laminin-521 or laminin-121 for nine passages. Variations in gene expression related to pluripotency, stemness, and male germ and somatic gonadal cells at different passages and different culture methods were evaluated. All cell lines expressed pluripotency markers at protein and gene level and were able to differentiate into cell types of the three germ layers after being cultured on laminin-521 for nine passages. Laminin 521 had no obvious effect on the expression of genes related to male germ cells and somatic gonadal cells when used as a matrix for hES cell cultures. Importantly, reduction in variation of pluripotency marker expression was observed after culturing the cells on laminin-521 compared to culture on hFFs, and could be reduced further with an increased culture period on laminin-521. Changing the cell culture medium of hES cells on hFFs from Dulbecco’s Modified Eagle Medium (DMEM) to NutriStem, which is commonly used for culturing the cells on matrices, did not affect the variation of pluripotency genes and genes related to stemness expression. hES cells cultured on laminin-521 were more homogenous, attached better and grew faster compared to hES cells cultured on matrigel. The results show that laminin-521 provides optimal culture conditions for adaptation to feeder and xeno-free conditions of hES cells derived and cultured on feeder cells. Moreover, laminin-521 has a positive effect on stabilizing and homogenizing pluripotent gene expression profiles between hES cell lines provides the first step towards more controllable and robust culture conditions for hES cells.