1. Human feeder layer system derived from umbilical cord stromal cells for human embryonic stem cells 
Meeyoung Cho, Ph.D., Eun Ju Lee, Ph.D., Hyun Nam, Ph.D., Ji-Hye Yang, M.S., Jaejin Cho, D.V.M., Ph.D., Jeong Mook Lim, D.V.M., Ph.D., Gene Lee, D.D.S., Ph.D. 
Fertility and Sterility 
Volume 93, Issue 8, Pages (May 2010)
DOI: /j.fertnstert
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Characterization of hUCSCs. (A) hUCSCs were subcultured every 3–4 days. (B) Flow cytometric analysis of hUCSCs indicated mesenchymal-like surface marker expression. Scale bar is 200 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Evaluation of pluripotency of human ESCs on hUCSCs. Pluripotency-specific markers of H9 cells on hUCSCs were analyzed. (A) H9 on MEFs were positive for Oct-4, SSEA-4, TRA-1-60, TRA-1-81, and ALP and were negative for SSEA-1. (B) H9 on hUCSCs at passage 30 were positive for Oct-4, SSEA-4, TRA-1-60, TRA-1-81, and ALP and were negative for SSEA-1. The expression of differentiation-related genes was detected in 4-week and 8-week EBs of H9 on MEFs and H9 on hUCSCs at passages 10 and 20. Scale bar = 200 μm. (C) H9 EBs cultivated on hUCSCs expressed three germ layer–related genes after spontaneous differentiation, as H9 EBs on MEFs did. Undifferentiated markers, Nanog and Oct-4; ectodermal marker, neurofilament 68 kDa; endodermal marker, α-fetoprotein; mesodermal marker, α-cardiac actin. Histologic analysis of the mass formed in SCID mice after injection of H9 cells on hUCSCs feeders after 30 passages exhibited teratocarcinoma. (D) A 12-week-old mass from H9 cells consisted of three germ layers: endoderm (epithelium), mesoderm (cartilage, bone, adipose tissue), and ectoderm (gut, goblet cell). Scale bar = 200 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

4. Figure 2
Evaluation of pluripotency of human ESCs on hUCSCs. Pluripotency-specific markers of H9 cells on hUCSCs were analyzed. (A) H9 on MEFs were positive for Oct-4, SSEA-4, TRA-1-60, TRA-1-81, and ALP and were negative for SSEA-1. (B) H9 on hUCSCs at passage 30 were positive for Oct-4, SSEA-4, TRA-1-60, TRA-1-81, and ALP and were negative for SSEA-1. The expression of differentiation-related genes was detected in 4-week and 8-week EBs of H9 on MEFs and H9 on hUCSCs at passages 10 and 20. Scale bar = 200 μm. (C) H9 EBs cultivated on hUCSCs expressed three germ layer–related genes after spontaneous differentiation, as H9 EBs on MEFs did. Undifferentiated markers, Nanog and Oct-4; ectodermal marker, neurofilament 68 kDa; endodermal marker, α-fetoprotein; mesodermal marker, α-cardiac actin. Histologic analysis of the mass formed in SCID mice after injection of H9 cells on hUCSCs feeders after 30 passages exhibited teratocarcinoma. (D) A 12-week-old mass from H9 cells consisted of three germ layers: endoderm (epithelium), mesoderm (cartilage, bone, adipose tissue), and ectoderm (gut, goblet cell). Scale bar = 200 μm.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

5. Figure 3
Karyotype analysis of human ESCs on hUCSCs. H9 cells on hUCSC feeder layers at 36 passages maintained a normal karyotype, as shown by G-band staining.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

6. Figure 4
bFGF secretion by hUCSCs. Early- (p5) and late-passage (p10) hUCSCs secreted bFGF at similar levels (P > 0.05), as detected by ELISA. The bFGF level in medium was decreased during 24 h incubation. Both early- and late-passage hUCSCs maintained higher bFGF secretion than did embryonic stem cells after 24 h incubation.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions