human embryonic stem cell line h7  (Geron Bio)

Journal: PLoS ONE

Article Title: A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells

doi: 10.1371/journal.pone.0161229

Figure Lengend Snippet: Characterization of hPSCs generated * and cultured in defined, xeno-free EOD hPSC medium. Percentage of cells expressing markers of pluripotency after cultivation or generation in the defined hPSC medium using flow cytometry; detection of ectoderm, mesoderm and endoderm markers after EB differentiation; results of G-banding, CGH/SNP analyses.

Article Snippet: Human embryonic stem cell (hESC) lines H7 (WA07 (MEF Platform), lot No. FTDL-01), H9 (WA09 (MEF Platform), lot No. DL-10) and H14 (WA14 (MEF Platform), lot No. DL-01) were obtained from WiCell (Madison, WI) and initially cultivated on mitomycin-C treated mouse embryonic fibroblasts (MEFs) in 80% Knockout-Dulbecco’s Modified Eagle’s Medium/F12 (Life Technologies, 12660–012), 20% Knockout Serum Replacement (Life Technologies, 10828–028), 2mM L-glutamine (Life Technologies, 25030–081), 55μM β-mercaptoetanol (Life Technologies, 21985–023), 1% non-essential amino acids (Life Technologies, 11140–050), and 8 ng/ml human basic fibroblast growth factor (Life Technologies, PHG0026).

Techniques: Generated, Cell Culture, Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells

doi: 10.1371/journal.pone.0161229

Figure Lengend Snippet: Human hESC line WA07 was serially subcultured for five passages. The number of viable cells per passage were compared by initially seeding 2×10 4 viable cells per cm 2 into three wells of a six-well plate for each medium, using respective passaging solution and matrix. Panel A shows cell attachment and growth of WA07 hESCs at passage 1 in L7 ™ hPSC, mTeSR1 TM , and E8 TM media. Panel B shows cell attachment and growth of WA07 hESCs at the end of passage 5 in L7 ™ hPSC, mTeSR1 ™ , and E8 ™ media. Panel C Panel A shows the viability and total viable cells of WA07 hESCs grown in L7 ™ hPSC, mTeSR1 ™ , and E8 ™ media, demonstrating comparative growth for the cells grown in each media. Panel D shows immunocytochemistry analysis of WA07 hESCs grown in L7 ™ hPSC, mTeSR1 ™ , and E8 ™ media, demonstrating comparative expression of OCT4 (red), Nanog (red), SSEA4 (green), TRA1-60 (green), and TRA1-81 (green) in each media. Following differentiation of hPSCs into embryoid bodies (Panel E), differentiated WA07 hESCs readily expressed the markers for early ectoderm (detected TUJ1, green), endoderm (detected Alpha-Feto Protein (AFP), green), and mesoderm (detected Smooth Muscle Actin (SMA), green) in L7 ™ hPSC and E8 TM media. Cell nuclei are shown by DAPI (blue).

Article Snippet: Human embryonic stem cell (hESC) lines H7 (WA07 (MEF Platform), lot No. FTDL-01), H9 (WA09 (MEF Platform), lot No. DL-10) and H14 (WA14 (MEF Platform), lot No. DL-01) were obtained from WiCell (Madison, WI) and initially cultivated on mitomycin-C treated mouse embryonic fibroblasts (MEFs) in 80% Knockout-Dulbecco’s Modified Eagle’s Medium/F12 (Life Technologies, 12660–012), 20% Knockout Serum Replacement (Life Technologies, 10828–028), 2mM L-glutamine (Life Technologies, 25030–081), 55μM β-mercaptoetanol (Life Technologies, 21985–023), 1% non-essential amino acids (Life Technologies, 11140–050), and 8 ng/ml human basic fibroblast growth factor (Life Technologies, PHG0026).

Techniques: Passaging, Cell Attachment Assay, Immunocytochemistry, Expressing