A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells

Abstract

We used successfully human foreskin fibroblasts as feeder cells for derivation and continued undifferentiated growth of hES cells. These feeder cells are convenient for IVF units, because no fetal human tissues or tissue from operations are needed.

Toward xeno-free culture of human embryonic stem cells

“…To eliminate the murine element from feeder cells, Hovatta et al (2003) have derived novel hESC lines on neonatal human foreskin fibroblasts. The hESC lines were grown on these fibroblasts for up to 9 months, at which time they still expressed appropriate undifferentiated hESC markers, had a normal karyotype and exhibited teratoma formation in SCID mice.…”

Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells

“…Thus, Diff Miz-hES6 feeder cells support the proliferation of hESCs as efficiently as MEFs. In contrast, the literature suggests hESCs cultured on other human feeder cells require a 5-7 day passaging interval (data not shown) (Richards et al, 2002;Amit et al, 2003a;Cheng et al, 2003;Hovatta et al, 2003;Lee et al, 2005). The hESC lines cultured on Diff Miz-hES6 feeder cells were relatively circular in shape (Figure 2), which is similar to their morphology when cultured on MEF feeder cells (data not shown).…”

“…Embryonic stem cells (ESCs) can differentiate into all the cells derived from three embryonic germ layers (Choi et al, 2004;Kim et al, 2005a;b). Human embryonic stem cells (hESCs) are derived from inner cell masses of human blastocysts (Thomson et al, 1998).…”

“…Recently, we and other groups demonstrated that it is possible to culture hESCs on feeder cells that originate from human sources (Richards et al, 2002;Amit et al, 2003a;Cheng et al, 2003;Hovatta Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells Lee et al, 2005). In most cases, however, these human feeder cell lines are derived from samples obtained from patients or discarded human fetuses.…”

Derivation of human embryonic stem cells at the Center of Regenerative Medicine in Barcelona

“…HFF were irradiated at 55 Gy and plated in 0.1% gelatine-coated IVF dishes (Falcon, Becton Dickinson, Franklin Lakes, NJ; 7 × 10 4 /cm 2 ; Hovatta et al, 2003). Whole blastocysts or isolated ICM's were transferred on feeder's dishes and cultured at 37°C and 5% CO 2 in KnockOut Dulbecco's modified Eagle's medium (KO-DMEM) supplemented with 2 mmol/l GlutaMAX (Gibco, Invitrogen), 0.05 mmol/l 2-mercaptoethanol (Gibco, Invitrogen), 8 ng/ml basic fibroblast growth factor (Invitrogen), 1% nonessential amino acids (Cambrex, East Rutherford, NJ), 20% KnockOut Serum Replacement (SR; Invitrogen), and 0.5% penicillin-streptomycin (Gibco, Invitrogen morphology (a compact colony structure, a high ratio of nucleus to cytoplasm, and prominent nucleoli) appeared among other cell types and unspecific growth.…”