Tumor Immune Cell Profiling by Flow Cytometry

Flow cytometry was carried out as previously described [11 (link)]. The mice were killed by cervical dislocation, and the tumor tissue was harvested, cut up, and incubated with 488 U/ml collagenase I (Sigma, #C0130), 230 U/ml collagenase Xi (Sigma, #C7657), 125 U/ml hyaluronidase (Sigma, #H3506), and 60 U/ml DNase I (Sigma, #d4527) at 37° C for 45 min. The digest was filtered by a 70 μm cell filter, and 1×10⁷ cells were stained with an FITC-conjugated F4/80 antibody from Abcam and a PE-conjugated CD206 antibody from R&D Systems and analyzed by flow cytometry.

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Chen Y., Tang G., Qian H., Chen J., Cheng B., Zhou C, & Shen Y. (2021). LncRNA LOC100129620 promotes osteosarcoma progression through regulating CDK6 expression, tumor angiogenesis, and macrophage polarization. Aging (Albany NY), 13(10), 14258-14276.

Publication 2021

collagenase I collagenase Xi hyaluronidase DNase I

Antibody Cells Cervical Collagenase Collagenase i Dislocation Dnase Fitc Flow cytometry Hyaluronidase Mice Tissue Tumor

Corresponding Organization :

Other organizations : Soochow University, Second Affiliated Hospital of Soochow University, Traditional Chinese Medicine Hospital of Kunshan, Shanghai University of Traditional Chinese Medicine, Yangzhou University

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Variable analysis

independent variables

Collagenase I (488 U/ml)
Collagenase XI (230 U/ml)
Hyaluronidase (125 U/ml)
DNase I (60 U/ml)

dependent variables

Percentage of F4/80-positive cells
Percentage of CD206-positive cells

control variables

Tumor tissue harvesting
Tissue digestion duration (45 min)
Cell filtering (70 μm)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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