Line name: ES[5]
Origin: embryonary

Data related to the biological sample:
Human embryo cryopreserved at day +2 of development
Frozen: 7.05.2001
Donated: 14.03.2006
Received at the Banco de líneas celulares of CMR[B]: 16.10.2006
Thawed: 16.10.2006

General description of the process:
The cryopreserved donated embryo was thawed with by a slow protocol with PROH and sucrose. After 4 days in culture, the zona pellucida (ZP) was removed by acid Tyrodes. 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) (Invitrogen), 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:
A cell clump with undifferentiated appearance was observed between the differentiated growth. After 15 days in culture, 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[5]

Karyotype ES[5]

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ES[5] (61,6 Kb)