CD expression and had been injected into NODSCID
CD expression and have been injected into NODSCID mice (4 mice per dose, two or extra doses per fraction). TICf was calculated as described in Fig. A. Matched samples are indicated by brackets inside the left margin. Samples in which all TICs had been located in the CD+ fraction are in boldface type. Samples in which all or most TICs were found within the CD- fraction are in italics. T, strong tumor; M, metastasis; A, ascites; p, xenograft passage number; blue type, recurrent disease; black variety, major disease; red type, principal disease treated with neo-adjuvant chemotherapy; ND, not determined. The total number of TICs in all fractions was calculated according to sorting cells as: (no. of CD+ or CD- cells per cells) (CD+ or CD- TICf). See Table S for calculation from the probability that tumors formed from CD- cells are as a consequence of contamination with CD+ cells (based on P .). NA, not applicable, as all TICs are in the CD+ or CD- fraction; YesNo, tumor formation from CD- cells is (or is not) explicable by contamination with CD+ cells (at the self-confidence level). In circumstances exactly where it was achievable to measure TICs in unfractionated, CD+ and CD- SOC cells, the recovery of TIC activity (i.etotal TICs from unfractionated cells compared with all the sum of CD+ and CD- TICs) ranged from toLow recovery could reflect losses for the duration of sorting, gating tactic or, potentially, a supportive effect of non-TICs on TICs; high recovery could indicate inhibitory effects of non-TICs on TICs. Also, the practical limitations around the quantity and range of tumor cell doses that may be tested plus the number of recipient mice per dose intrinsically limit the accuracy of LDAs.Key Tumors Xenogra sTICs. Related events could explain the coexistence of CD+ and CD- TICs in other SOCs. Moreover, an analogous switch in CSC phenotype is believed to take place during conversion in the chronic phase of chronic myeloid leukemia, in which CSCs reside inside the hematopoietic stem cell compartment, to myeloid blast crisis where CSCs arise from granulocytemacrophage progenitorsIt will probably be intriguing to see if certain genetic alterations correlate with the distinct TIC phenotypes; certainly, preliminary copy number variation analyses reveal dramatic variations in xenografts derived from some CD- TICs compared with parental CD+ TICs. Alternatively, given that CD clearly will not be an obligate, functional marker of SOC TICs, we cannot exclude the possibility that the CD- TICs that arise in xenografts basically reflect the influence in the altered microenvironment on CD expression. Regardless, the increased number and frequency of CD- TICs, in combination using the observed histological adjustments in early passage xenografts, argue for intense caution when utilizing these models to infer the phenotype of patient TICs. Moreover, the phenotypic heterogeneity and instability of TICs could OICR-9429 web complicate efforts to translate CSC concepts towards the clinic. Nonetheless, our identification of distinctive phenotypic classes of TICs in major patient samples gives a platform for the identification of additional selective markers and also the detailed .orgcgidoi..molecular characterization of this PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22072678?dopt=Abstract potentially critical population of SOC cells. Components and MethodsHigh-grade SOC samples had been obtained from the University 
Health Network Tissue Bank with patient consent. Samples were mechanicallyenzymatically digested to generate single-cell suspensions and red blood cellsleukocytes were depleted. Flow cytometry and FACS have been carried out as de.