cd82 apc  (Miltenyi Biotec)

Journal: Cells

Article Title: Long-Term Engraftment and Satellite Cell Expansion from Human PSC Teratoma-Derived Myogenic Progenitors

doi: 10.3390/cells14151150

Figure Lengend Snippet: Scheme outlining the key steps of the study. hiPSCs were injected into TA muscles of NSG mice, and teratomas were harvested two months later. Tissue was digested, and skeletal myogenic progenitor cells co-expressing CD82, ERBB3, and NGFR were sorted by FACS, cultured in vitro, tested for differentiation into myotubes, and, in parallel, transplanted into tibialis anterior (TA) muscles of injured NSG-mdx 4Cv mice following X-ray and cardiotoxin injection. TA muscle tissue was harvested at various time points, and stained sections were analyzed with different antibodies to assess human cell contribution, muscle regeneration, and fiber type differentiation.

Article Snippet: The dissociated cells were incubated with antibodies APC anti-CD82 (FAB4616A, R&D Systems, Minneapolis, MN, USA; RRID: AB_2076404), PE anti-ERBB3 (304706, BioLegend, San Diego, CA, USA; RRID: AB_2099569), and PECy7 anti-NGFR (Cat# 562122, BD Biosciences, San Jose, CA, USA; RRID: AB_10894762); each at 0.5 uL per 1 million cells) in FACS buffer for one hour on ice.

Techniques: Injection, Muscles, Expressing, Cell Culture, In Vitro, Staining

Journal: Cells

Article Title: Long-Term Engraftment and Satellite Cell Expansion from Human PSC Teratoma-Derived Myogenic Progenitors

doi: 10.3390/cells14151150

Figure Lengend Snippet: ( A ) Morphology of teratomas formed in TA muscles of NSG mice two months after injection of hiPSCs. ( B ) Isolation of skeletal myogenic progenitor cells co-expressing CD82, ERBB3, and NGFR by FACS from TA teratomas. ( C ) Expression levels of various myogenic and fibroblastic markers under different growth factor conditions, as determined by RNA sequencing, presenting mean ratio normalized counts. ( D ) Morphology of isolated skeletal myogenic cells cultured at different passages. PAX7 staining of cells in the proliferative phase at confluency, from the same passage used for transplantation. ( E ) MyoD, Myogenin, MF-20, and PAX7 staining of teratoma-derived skeletal myogenic cells in differentiation medium at passage 3, highlighting their capacity to form myotubes. ( F ) Percentage of cells positive for PAX7 in proliferative and differentiation media, and for MyoD, Myogenin, and MF-20 in differentiation medium. Data were obtained from five fields per sample across three replicates.

Article Snippet: The dissociated cells were incubated with antibodies APC anti-CD82 (FAB4616A, R&D Systems, Minneapolis, MN, USA; RRID: AB_2076404), PE anti-ERBB3 (304706, BioLegend, San Diego, CA, USA; RRID: AB_2099569), and PECy7 anti-NGFR (Cat# 562122, BD Biosciences, San Jose, CA, USA; RRID: AB_10894762); each at 0.5 uL per 1 million cells) in FACS buffer for one hour on ice.

Techniques: Muscles, Injection, Isolation, Expressing, RNA Sequencing, Cell Culture, Staining, Transplantation Assay, Derivative Assay