Journal: Reproductive biomedicine online

Article Title: Improved cryopreservation of human embryonic stem cells with trehalose.

doi: 10.1016/s1472-6483(10)61692-6

Figure Lengend Snippet: Figure 2. Characterization of human embryonic stem (hES)1 cells at passage 38. (A) Alkaline phosphatase staining of hES1 colonies. (B–G) Immunofl uorescent staining of hES1 colonies with monoclonal antibodies against stage-specifi c embryonic antigens SSEA-3 (B), SSEA-4 (C), tumour rejection antigens TRA-1–60 (D), TRA-1–81 (E), SSEA-1 (F), and octamer-binding transcription factor Oct-4 (G). (H) Reverse transcription-polymerase chain reaction (RT-PCR) analysis of Oct-4 expression: Lane 1, DNA ladder (100 bp); lane 2, human embryonic fi broblast (hEF) cells as negative control; lanes 3–5, hES1 cells at passage 13, 26 and 38 respectively; lane 6, terato-2 cells as positive control. (I) Telomerase activities analysed by Telo/PCR-enzyme-linked immunosorbent assay kit. hEF and 293 cells were used as negative and positive controls, respectively. Scale bars: 200 μm.

Article Snippet: The medium consisted of knockout Dulbecco’s modifi ed Eagle’s medium (KO-DMEM; Invitrogen, Carlsbad, CA, USA), supplemented with 20% knockout serum replacement (Invitrogen) or ES culture grade fetal bovine serum (Hyclone, Logan, UT, USA), 1% non-essential amino acid, 50 IU/ml penicillin and 50 μg/ml streptomycin, 2 mmol/l L-glutamine, 0.1 mmol/l β-mercaptoethanol (all from Invitrogen), and 4 ng/ml human recombinant β-fi broblast growth factor (R and D Systems, Minneapolis, MN, USA).

Techniques: Staining, Bioprocessing, Binding Assay, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Negative Control, Positive Control, Enzyme-linked Immunosorbent Assay