Human Embryonic Stem Cell Marker Panel (Hicks et al., 2020 (link); Roth et al., 2020 (link); Bomba et al., 2021 (link); Osnato et al., 2021 (link)) (Abcam, ab238602) was used to analyze pluripotency protein markers by immunofluorescence. A Trilineage Differentiation Kit (Ward et al., 2017 (link)) (STEMCELL, #05230) was used to induce differentiation into the three germ layers. Human three Germ Layer 3-Color Immunocytochemistry Kit (Han et al., 2020 (link)) (R&D, SC022) was used to identify specific protein markers of the three germ layers. A Giemsa Stain solution (Solarbio, G1015) was used to analyze the karyotype. Fluorescence images were obtained under an LSM900 laser scanning confocal microscope (Zeiss, Germany).
Trilineage differentiation kit
Manufactured by STEMCELL
Sourced in Canada
The Trilineage differentiation kit is a laboratory tool designed for the in vitro differentiation of stem cells into the three primary germ layers: endoderm, mesoderm, and ectoderm. The kit provides a standardized protocol and optimized media formulations to facilitate the directed differentiation of stem cells towards specific cell lineages.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 900 confocal laser scanning microscope, by Zeiss (1 mentions)
Human three germ layer 3 color immunocytochemistry kit, by R&D Systems (1 mentions)
Giemsa stain solution, by Solarbio (1 mentions)
Mycoalert kit, by Lonza (1 mentions)
Cytotune2.0 ips sendai reprogramming kit, by Thermo Fisher Scientific (1 mentions)
6 protocols using trilineage differentiation kit
1
Comprehensive Stem Cell Characterization
2
Trilineage Differentiation and RPE Induction
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Trilineage differentiation assays were carried out according to instructions of the Trilineage differentiation kit (Stem Cell Technology). Briefly, appropriate amounts of ES cells were seeded on Matrigel-coated plates. Then, cells were treated with lineage-specific differentiation medium respectively. About 5 or 7 days later, typical germ layer markers were detected by immunostaining. The antibodies used are listed in Table 2 .
For directed differentiation of LRP2-KO HESC cells into RPE, a previously published protocol was used with mild modification (Foltz and Clegg, 2017 ) and cytokines were added step by step to induce stem cell transformation.
For directed differentiation of LRP2-KO HESC cells into RPE, a previously published protocol was used with mild modification (Foltz and Clegg, 2017 ) and cytokines were added step by step to induce stem cell transformation.
3
Generating patient-specific iPSCs
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iPSCs were generated reprogramming fibroblasts from healthy controls and CMT2B patients carrying the RAB7V162M mutation by Sendai viral transduction of the transcription factors OCT4, SOX2, KLF4, and c-MYC (CytoTune2.0-iPS Sendai Reprogramming Kit, Thermo Fisher, Waltham, MS, USA). iPSCs were then tested for Sendai virus clearance (CytoTune2.0-iPS Sendai Reprogramming Kit, Thermo Fisher) and pluripotency (Trilineage differentiation Kit, Stem Cell Technologies, Vancouver, Canada). Cells were regularly screened and confirmed negative for mycoplasma during both maintenance and differentiation (MycoAlert kit, Lonza, Basel, Switzerland).
4
Tri-Lineage Embryoid Body Differentiation
5
Tri-Lineage Embryoid Body Differentiation
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Embryoid bodies were generated using a tri-lineage differentiation kit (Stem Cell Technologies). Germ layer differentiation was assessed by immunochemistry.
6
Trilineage Differentiation and RPE Induction
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