Available Lines > Characterization of the ES2 Cell Line

Line name: ES[2]
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) (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:
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[2]

Code

ES[2]

 

 

Embryo origin

FIV Institut Dexeus

 

 

Characterization

 

 

 

Passage nº

21

 

 

Feeders

Human foreskin Fibroblasts

 

 

ICM isolation

no

 

 

Karyotype

46, XY

 

 

Phenotype

 

SSEA-1

-

SSEA-3

+

SSEA-4

+

TRA1-60

+

TRA1-81

+

Oct4

+

Sox 2

+

Nanog

+

Alkaline Phosphatase

+

 

 

Freeze/thaw viability

si

 

 

Pluripotency

 

In vivo

si

In Vitro:
   ectoderm (β-tubulin III)
   endoderm (α-fetoprotein)   
   mesoderm (myosine)

 

+
+
+

 

 

Microbiologic analisys

 

Aerobics

-

Anaerobics

-

Fungus

-

Mycoplasma

-

 

 

HLA

HLA-A*0101
HLA-B*0801
HLA-Cw*0701
HLA-DRB1*0301
HLA-DQB1*0201

 

 

fingerprinting

done

Karyotype ES[2]


PDF document

ES[2] (62,2 Kb)



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