The engineered cell sheets were fixed in 10% neutral buffered formalin and embedded in paraffin. The tissue sections were used for immunofluorescent staining with CD19 (Cat # NBP2-25196), CD73 (Cat # NBP2-25237) (NovusBIo, CO, United States), CD29 (Cat # ab134179), HLA-A (Cat # ab52922), HLA-DR (Cat # ab92511) (Abcam, MA, United States), and Oct3/4 (Cat # NB-100-2379SS) (Novus Biologicals LLC., Littleton, CO, United States). Alexa Fluor 488 donkey anti-rabbit conjugated secondary antibodies and Alexa Fluor 488 donkey anti-mouse conjugated secondary antibodies (Invitrogen, Carlsbad, CA, United States) were used. Propidium iodide (Invitrogen, Carlsbad, CA, United States) was used to stain nuclear DNA. An EVOS M5000 microscope was used to analyze the slides (Invitrogen, Carlsbad, CA, United States).
Hla a
Manufactured by Abcam
Sourced in United States
HLA-A is a class I major histocompatibility complex (MHC) protein. It plays a central role in the immune system by presenting peptide fragments to T-cell receptors. The HLA-A gene is highly polymorphic, with many different alleles identified in the human population.
Automatically generated - may contain errors
Lab products found in correlation
Hla dr, by Abcam (3 mentions)
Propidium iodide, by Thermo Fisher Scientific (2 mentions)
Evos m5000 microscope, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 donkey anti rabbit conjugated secondary antibodies, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 donkey anti mouse conjugated secondary antibodies, by Thermo Fisher Scientific (2 mentions)
Traf1, by Abcam (1 mentions)
Traf2, by Abcam (1 mentions)
Propidium iodine, by Thermo Fisher Scientific (1 mentions)
Col 2, by Abcam (1 mentions)
Alexa fluor 488 donkey anti rabbit conjugated second antibodies, by Thermo Fisher Scientific (1 mentions)
Bglap, by Thermo Fisher Scientific (1 mentions)
Srebp1, by Thermo Fisher Scientific (1 mentions)
Pparg, by Cell Signaling Technology (1 mentions)
Cd105, by Thermo Fisher Scientific (1 mentions)
5 protocols using hla a
1
Phenotypic Characterization of Engineered Cell Sheets
2
Immunohistochemical Profiling of Biomarkers
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Serial 4-μm-thick sections of fixed tissue were cut and placed onto glass slides, after which they underwent IHC staining with primary polyclonal antibodies targeting EBV-LMP1, EBNA1, ACOT1, ACOT4, ALCAM, BRAF, PDL1, B7H3, CD4, CD87, CXCL12, CXCL13, fatty acid desaturase (FADS) 2, FADS3, FADS6, hydroxyacyl-CoA dehydrogenase (HADHA), human leukocyte antigens (HLAA), human leukocyte antigens B (HLAB), human leukocyte antigens C (HLAC), PPT1, PPT2, TRAF1, and TRAF2 (all primary antibodies were purchased from Abcam, Hong Kong, China, and are listed in the Supplemental Digital Content 1 [see Table S1, http://links.lww.com/CTG/A87 ]). The sections were washed and incubated with an amplification agent and a polymerase. Nuclei were counterstained with hematoxylin. The expression of proteins was scored by 2 pathologists who were blinded and were unaware of all clinical parameters. Where serious discrepancies arose, a final score was determined by reassessment of the staining using a multihead microscope.
3
Characterization of Engineered Cell Sheets
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Engineered cell sheets were fixed in 10% neutral buffered formalin and embedded in paraffin. Tissue sections were used for immunofluorescent staining with CD19, CD73 (NovusBIo, CO, USA), CD29, HLA-A, HLA-DR (Abcam, MA, USA), Oct3/4 (Novus Biologicals LLC., Littleton, CO, USA). Alexa Fluor 488 donkey anti-rabbit conjugated secondary antibodies and Alexa Fluor 488 donkey anti-mouse conjugated secondary antibodies (Invitrogen, Carlsbad, CA, USA) were used. Propidium iodide (Invitrogen, Carlsbad, CA, USA) was used to stain nuclear DNA. A EVOS M5000 microscope was used to analyze the slides (Invitrogen, Carlsbad, CA, USA).
4
Histological Analysis of Mouse Tissues
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For histological analysis, formalin-fixed mouse tissues were embedded in paraffin, sectioned, stained with hematoxylin and eosin (H&E), CD3 (Serotec), or HLA-A (Abcam) for immunohistochemistry (IHC) and imaged by a Pannoramic 250 scanner.
5
Immunofluorescent Characterization of Cell Sheets
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Engineered cell sheets were fixed in 10% neutral buffered formalin and embedded in paraffin. Tissue sections were then stained with hematoxylin & eosin (H&E) or used for immunofluorescent staining with CD19, CD73, Acetyl-CoA1 (NovusBIo, CO, USA), CD29, COLII (Collagen II), HLA-A, HLA-DR (Abcam, MA, USA), CD45R, CD105, BGLAP (Osteocalcin), SREBP1 (ThermoFisher Scientific, CA, USA), PPARg (Cell Signaling, MA, USA), ACAN (Aggrecan) and SPARC (ThermoFisher Scientific). Alexa Fluor R 488 donkey antimouse conjugated second antibodies, Alexa Fluor 488 donkey antirabbit conjugated second antibodies and Alexa Fluor 488 donkey antirat conjugated second antibodies (Invitrogen, OR, USA) were used. Propidium iodine (Invitrogen) was used to stain nuclear DNA. A Nikon 400 fluorescence microscope was used to analyze the slides (Nikon Inc., NY, USA).
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