Line name: ES[3]
Origin: embryonary

Data related to the biological sample:
Human embryo cryopreserved at day +6 of development (blastocyst)
Frozen: 14.06.1999
Donated: 7.03.2006
Received at the Banco de líneas celulares of CMR[B]: 9.05.2006
Thawed: 9.05.2006

General description of the process:
The cryopreserved donated embryo was thawed with by a slow protocol with PROH and sucrose. After 24h in culture, the zona pellucida (ZP) was removed by pronase. The dezoned blastocyst was cultured over a monolayer of irradiated fibroblasts in hES culture medium.

Cell and culture medium used for derivation:
Cell: human foreskin fibroblasts (ATCC, American Type Culture Collection, CCD1112Sk).
Culture medium: Knockout Dulbecco?s modified Eagle?s medium supplemented with 2 mmol/l GlutaMAX (Gibco, InVitrogen corporation), 0,05mmol/l 2-mercaptoethanol (Gibco, InVitrogen corporation), 8 ng/ml basic fibroblast growth factor (bFGF) (InvitKaryotype*rogen), 1% non-essential amino acids (Cambrex), 20% Knockout Serum Replacement (InVitrogen) y 0,5% Penicillin-Streptomycin (Gibco, InVitrogen corporation).

General description of derivation and cell line maintenance:
After 12 days in culture a cell clump appeared. It was mechanically dissociated and re-plated on a new monolayer of fibroblasts. The cells are passaged every 6-7 days. The passaging is done mechanically.

Freezing of the colonies is done by a slow protocol with 90% FBS (fetal bovine serum) and 10% DMSO (dimethylsulphoxide).

Characterization ES[3]

Karyotype ES[3]

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ES[3] (61,6 Kb)