ISCI/FRM/004 – hES Cell Details
Please complete a form for each hES cell line
PHOTOCOPY ADDITIONAL FORMS, AS REQUIRED
Participating Laboratory:
Contact
Name:
L. Ahrlund-Richter
Karolinska Institute
Stockholm, Sweden
Marta Imreh
Dept of Lab. Med.
Karolinska Institute
141 57 Stockholm, Sweden
E-mail:
marta.imreh@ki.se
Tel:
+ 46 8 5858 3641
Cell Line: HS181
CELL LINE DERIVATION:
Embryo details
4BB
Embryo used (please tick)
Fresh

Frozen
Was the embryo known to carry any mutations?
Yes

No
If so, please provide details:
Isolated Inner Cell Mass
Was the line derived from whole embryo or isolated inner cell mass (please tick)?
ICM isolated by
Whole embryo
ICM isolated by
Mechanical dissociation
immunosurgery

If immunosurgery, what antibody and complement was used?
Rabbit anti Human whole serum (Sigma)
Guinea pig complement serum (Sigma)
Media used (give details)
Knockout D-MEM 13 (GibcoBRL, Life
Technologies), supplemented with 2 nmol/l Lglutamine, 20% FCS (R&D, Sweden), 0.1 mmol/l
b-mercaptoethanol (Gibco), 1% non-essential amino
acids (Gibco), and recombinant human LIF, 1 ml/ml
(Chemicon, UK).
Feeder cell used (give details
Human foreskin fibroblast line; CRL2429 (ATCC)
Time to first passage:
1w
Subculture method used for first passage:
Mechanical
SUBSEQUENT CELL LINE MAINTENANCE
Feeder cells used (give details):
Litt. ref: Imreh, P., Wolbank, S., Unger, C., Gertow, K., Aints,
A., Szeles, A., Imreh, S., Hovatta, O., Fried, G., Dilber, S., and
Ährlund-Richter, L. Culture and expansion of the human
embryonic stem cell line HS181, evaluated in a double color
system. Stem Cells and Development Vol. 13:337-343, 2004
The HS181 cells were cultured in 80% KO-Dulbecco’s
modified Eagle medium (DMEM) (Invitrogen/Gibco-BRL;
PL 10829018), 20% serum replacement/SR
(Invitrogen/Gibco-BRL; PL10828028), 2 mM L-glutamine
(Invitrogen/Gibco-BRL; 25030024), 1% nonessential
amino acids (Invitrogen/Gibco-BRL; 11140035), 0.1 mM
beta-mercapoethanol (Invitrogen/Gibco-BRL; 31350010),
and 4 ng/ml fibroblast growth factor (bFGF)
(Invitrogen/Gibco-BRL; 13256029), Penic./Strept.
(Invitrogen/Gibco-BRL; PL 15140122).
Human foreskin fibroblast line; CRL2429 (ATCC)
Population doubling time, if known
24-36 h
Subculture protocol used (give details):
Media used (give details):
Karyotype of cells – please include passage level(s) at which karyotyping was performed
46XX at passages 22, 32, 39, 45, 54
Has there been any alteration over time in:
YES
NO
1
(please tick)
X
Culture conditions
X
Cell Characteristics
2
X
Karyotype
X
Differentiation
X
Other
If YES, please provide details:
1
Derived using FCS, now passaged using SR
2
Karyotype changes; trisomy 12
(Imreh, M, Gertow, K, Cedervall, J, Unger, C, Holmberg, C, Szöke, K, Csöregh, L, Fried, G, Dilber, S,
Blennow, E, and Ährlund-Richter, L. “In vitro culture conditions favoring selection of chromosomal
abnormalities in human ES cells”. Journal of Cellular Biochemistry 1;99(2):508-16. 2006)
Any other comments/information that you think would be useful to this project
HS181 at passage numbers 28/29 (time point 1) and 33/34 (time point 2) were used in the current project.
Litt.ref. on derivation of HS181.
Hovatta, O, Mikkola, M, Gertow, K, Strömberg, A-M, Inzunza, J, Hreinsson, J, Rozell, B, Blennow, E.,
Andäng, M., Ährlund-Richter, L. A culture system using human foreskin fibroblasts as feeder cells allows
production of human embryonic stem cells.
Human Reproduction 18 (7): 1404-1409, 2003
PLEASE CONTINUE ON ANOTHER SHEET OF PAPER, IF REQUIRED