A Humanized Stromal Bed Is Required for Engraftment of Isolated Human Primary Squamous Cell Carcinoma Cells in Immunocompromised Mice  Girish K. Patel,

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A Humanized Stromal Bed Is Required for Engraftment of Isolated Human Primary Squamous Cell Carcinoma Cells in Immunocompromised Mice  Girish K. Patel, Carole L. Yee, Stuart H. Yuspa, Jonathan C. Vogel  Journal of Investigative Dermatology  Volume 132, Issue 2, Pages 284-290 (February 2012) DOI: 10.1038/jid.2011.284 Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Creation of a squamous cell carcinoma (SCCa) in vivo xenograft model that can accurately recapitulate and propagate human SCCa from intact tumor tissue. (a) Successful engraftment of primary human SCCa tissue required before creation of a stromal bed, into which the tissue is implanted after 14 days. After 12 weeks, tumor growth was determined by histological sampling of all xenograft sites. (b) Hematoxylin and eosin-stained sections of athymic nude mouse skin demonstrate a fibrovascular proliferation following subcutaneous implantation of either a glass disc or Gelfoam dressing for 2 weeks. (c) Tumor growth after implantation of intact SCCa tissue of similar size. (d) Xenograft tumors that developed from intact, primary human SCCa preserved the original tumor morphology. Bar=100μm. Journal of Investigative Dermatology 2012 132, 284-290DOI: (10.1038/jid.2011.284) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Xenografts express human markers and can be passaged in vivo. Primary human squamous cell carcinoma (SCCa)-derived xenograft tumors expressed human (a) HLA antigen, (b) cytokeratin, and (c) nuclear accumulation of mutant p53. Respective isotype controls are shown in panels underneath. Xenograft tumor stroma was predominantly replaced by murine cells that expressed murine major histocompatibility complex (MHC) class 1. Nonspecific staining was observed within the dead squamous accumulations. (d) Recreation of the original tumor morphology upon in vivo serial passage of the previously xenografted tumor. Histology after 12 weeks was similar to the original and primary xenograft tumors. Bar=100μm. Journal of Investigative Dermatology 2012 132, 284-290DOI: (10.1038/jid.2011.284) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Squamous cell carcinoma (SCCa) mechanical dissociation and generation of a single-cell suspension. (a) Primary human SCCa was (b) mechanically dissociated using surgical scissors, and (c) the minced tumor tissue was then implanted onto wound beds created by either a glass disc or Gelfoam dressing that had been implanted for 14 days in the athymic nude mice. Shown is a typical tumor after 12 weeks. (d) Cell suspensions were created by additional enzymatic dissociation so that few cells remained in the remnant tissue. (e) Tumor cells could be labeled with a human pancytokeratin antibody and (f) approximately 70% of the tumor-dissociated cells were viable, as assessed by FACS. Bars=100μm. Journal of Investigative Dermatology 2012 132, 284-290DOI: (10.1038/jid.2011.284) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Xenografts can accurately recapitulate and propagate human squamous cell carcinoma (SCCa) from cell suspensions. (a) Schematic. (b) SCCa xenografts from cell suspensions recreated the original tumor morphology (c) with no difference in xenograft growth based on the original tumor histological grade. Xenograft success was only dependent upon the number of tumor cells implanted and (d) harvested tumor tissue could be dissociated and successfully implanted into naive athymic nude mice. Bar=100μm. Journal of Investigative Dermatology 2012 132, 284-290DOI: (10.1038/jid.2011.284) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions