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. 2010 Apr;46(3-4):337-44.
doi: 10.1007/s11626-010-9308-0. Epub 2010 Mar 3.

Derivation of 30 human embryonic stem cell lines--improving the quality

Susanne Ström  1 , Frida Holm, Rosita Bergström, Anne-Marie Strömberg, Outi Hovatta
Affiliations

Derivation of 30 human embryonic stem cell lines--improving the quality

Susanne Ström et al. In Vitro Cell Dev Biol Anim. 2010 Apr.

Abstract

We have derived 30 human embryonic stem cell lines from supernumerary blastocysts in our laboratory. During the derivation process, we have studied new and safe method to establish good quality lines. All our human embryonic stem cell lines have been derived using human foreskin fibroblasts as feeder cells. The 26 more recent lines were derived in a medium containing serum replacement instead of fetal calf serum. Mechanical isolation of the inner cell mass using flexible metal needles was used in deriving the 10 latest lines. The lines are karyotypically normal, but culture adaptation in two lines has been observed. Our human embryonic stem cell lines are banked, and they are available for researchers.

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Figures

Figure 1
Figure 1
Derivation of the hESC line HS475. (a) Hatched blastocyst at Day 7 after fertilization, just before mechanical isolation of the ICM. (b) The ICM after derivation on human feeder layer. (c) Outgrowth of the ICM, 5 d after derivation. (d) HS475 at passage level 1. (e) HS475 at passage level 5. Magnification: (a) 200x (b, c) 400x (d, e) 40x original magnification
Figure 2
Figure 2
Typical morphology of hESC colonies. (a) HS181 on Matrigel, 40 x original magnification (b) HS401 on Matrigel, 400 x (c) HS366 on hff, 40 x. (d) HS420 on hff, 400 x.
Figure 3
Figure 3
Characterization of hESC lines by immunocytochemistry. (a) HS401 p37 Positive staining for nuclear marker Nanog (b) 366 p35 staining for nuclear marker Oct 3/4 (c) 362 p36 staining for TRA1–81 (d) 362 p36 staining for TRA-1–60. Magnification (a, c) 100× (b, d) 200× original magnification
See this image and copyright information in PMC

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