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
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