ABSTRACT
Background: Obstructive sleep apnea (OSA) is a chronic disorder of the upper airway. OSA surgery has oftentimes been researched based on the outcomes of single-institutional facilities. We retrospectively analyzed a multi-institutional national database to investigate the outcomes of OSA surgery and identify risk factors for complications. Methods: We reviewed the American College of Surgeons National Surgical Quality Improvement Program (NSQIP) database (2008−2020) to identify patients who underwent OSA surgery. The postoperative outcomes of interest included 30-day surgical and medical complications, reoperation, readmission, and mortality. Additionally, we assessed risk-associated factors for complications, including comorbidities and preoperative blood values. Results: The study population included 4662 patients. Obesity (n = 2909; 63%) and hypertension (n = 1435; 31%) were the most frequent comorbidities. While two (0.04%) deaths were reported within the 30-day postoperative period, the total complication rate was 6.3% (n = 292). Increased BMI (p = 0.01), male sex (p = 0.03), history of diabetes (p = 0.002), hypertension requiring treatment (p = 0.03), inpatient setting (p < 0.0001), and American Society of Anesthesiology (ASA) physical status classification scores ≥ 4 (p < 0.0001) were identified as risk-associated factors for any postoperative complications. Increased alkaline phosphatase (ALP) was identified as a risk-associated factor for the occurrence of any complications (p = 0.02) and medical complications (p = 0.001). Conclusions: OSA surgery outcomes were analyzed at the national level, with complications shown to depend on AP levels, male gender, extreme BMI, and diabetes mellitus. While OSA surgery has demonstrated an overall positive safety profile, the implementation of these novel risk-associated variables into the perioperative workflow may further enhance patient care.
ABSTRACT
BACKGROUND: The aim of the present study was to examine the cytostatic effects of cold atmospheric plasma (CAP) on different head and neck squamous carcinoma (HNSCC) cell lines either in isolation or in combination with low dose cisplatin. The effect of CAP treatment was investigated by using three different HNSCC cell lines (chemo-resistant Cal 27, chemo-sensitive FaDu and OSC 19). MATERIALS AND METHOD: Cell lines were exposed to CAP treatment for 30, 60, 90, 120 and 180 s (s). Cisplatin was added concurrently (cc) or 24 h after CAP application (cs). Cell viability, DNA damage and apoptosis was evaluated by dye exclusion, MTT, alkaline microgel electrophoresis assay and Annexin V-Fit-C/PI respectively. RESULTS: In all cell lines, 120 s of CAP exposure resulted in a significant reduction of cell viability. DNA damage significantly increased after 60 s. Combined treatment of cells with CAP and low dose cisplatin showed additive effects. A possible sensitivity to cisplatin could be restored in Cal 27 cells by CAP application. CONCLUSION: CAP shows strong cytostatic effects in HNSCC cell lines that can be increased by concurrent cisplatin treatment, suggesting that CAP may enhance the therapeutic efficacy of low dose cisplatin.
Subject(s)
Carcinoma, Squamous Cell , Cytostatic Agents , Head and Neck Neoplasms , Plasma Gases , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Cisplatin/pharmacology , Cisplatin/therapeutic use , Cytostatic Agents/therapeutic use , Head and Neck Neoplasms/drug therapy , Humans , Plasma Gases/pharmacology , Plasma Gases/therapeutic use , Squamous Cell Carcinoma of Head and Neck/drug therapyABSTRACT
OBJECTIVE: Ear, nose and throat infections are among the most common reasons for absence from work. They are usually caused by various bacteria like Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes. Cold atmospheric plasma (CAP) can effectively eliminate even multi-resistant bacteria and has no cytotoxic or mutagenic effects on the mucosa when applied for less than 60s. Aim of the study was to evaluate the effects of CAP on common ENT bacteria and on the mucosa of the upper aerodigestive tract. METHODS: The bactericidal effects of CAP against the bacteria most commonly causing ENT infections were investigated using the colony-forming units assay (CFU) on a Müller-Hinton agar plate after applying CAP for 30, 60, 90 and 120s. To evaluate the interaction of CAP with mucosal cells, 3D mini organ cultures were treated for up to 180s, after which cell viability and necrosis induction were evaluated. RESULTS: Treatment with CAP for 60s or longer induced at least a 3-log10 reduction in the bacterial load (> 99.9%). Treatment times shorter than 60s had only slight cytotoxic effects on cell viability and necrosis whereas treatment times above 60s showed a fast increase of cytotoxic side effects. CONCLUSION: CAP exhibited strong bactericidal effects on the most common ENT pathogens. Treatment times of up to 60s showed only minimal adverse reactions in healthy mucosa. CAP could be a promising new therapeutic modality for ENT infections.
Subject(s)
Haemophilus influenzae/drug effects , Otitis/microbiology , Plasma Gases/pharmacology , Respiratory Mucosa/drug effects , Respiratory Tract Infections/microbiology , Staphylococcus aureus/drug effects , Streptococcus pneumoniae/drug effects , Streptococcus pyogenes/drug effects , Adult , Cell Survival/drug effects , Colony Count, Microbial , Female , Humans , Male , Middle Aged , Necrosis , Organ Culture Techniques , Oropharynx , Pharyngitis/microbiology , Respiratory Mucosa/pathology , Rhinitis/microbiology , Young AdultABSTRACT
BACKGROUND: The surgeon's evaluation of resection status based on frozen section analysis during operation and pathological examination of resected specimens often differ. For this study, we recapitulated the surgeon's perspective during an operation, accordingly classified the evaluation of margins by the surgeon, and analyzed its impact on the outcome compared with the pathological results. METHODS: This was a retrospective analysis. As data sources, paper-based and digital patient files, as well as the Munich Cancer Registry database were used. RESULTS: Three hundred ninety-six cases were included in this analysis. Only the evaluation of margins by the surgeon influenced local control, and the pathological results influenced disease-free survival (DFS). Surprisingly, margins of >5 mm of normal tissue to cancer growth led to local control and overall survival (OS) significantly worse than 1 to 5-mm resections. CONCLUSION: The evaluation of margins by the surgeon is of significant importance for local control and OS. It is largely based on frozen section analysis, which, therefore, should be used whenever possible.
Subject(s)
Margins of Excision , Neoplasm Recurrence, Local/epidemiology , Squamous Cell Carcinoma of Head and Neck/pathology , Squamous Cell Carcinoma of Head and Neck/surgery , Adult , Aged , Aged, 80 and over , Disease-Free Survival , Female , Frozen Sections , Humans , Male , Middle Aged , Neoplasm Recurrence, Local/pathology , Retrospective Studies , Squamous Cell Carcinoma of Head and Neck/mortality , Survival Rate , Young AdultABSTRACT
The decision whether to perform an elective neck dissection in patients with head and neck squamous cell carcinoma and clinically negative lymph nodes (cN0) is made based on the probability of micrometastases in the neck for the given subsite and size of the primary. A retrospective chart review was performed of 203 patients with hypopharyngeal SCC who received a bilateral neck dissection. The frequency of histologically unveiled bilateral neck metastases was determined. A high frequency of contralateral metastases above 20% was detected for all carcinomas affecting the midline and those involving the medial wall of the pyriform sinus except of T1-stages (13%) and a low frequency for laterally located primaries (3%). Ipsilateral nodal status predicted contralateral neck metastases. Bilateral neck dissection should be recommended for primaries affecting the midline and T2-4 tumors involving the medial wall of the pyriform sinus.
Subject(s)
Carcinoma, Squamous Cell/secondary , Hypopharyngeal Neoplasms/pathology , Neck Dissection , Neoplasm Micrometastasis/diagnosis , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/pathology , Female , Humans , Hypopharynx/pathology , Lymphatic Metastasis/diagnosis , Male , Middle Aged , Probability , Retrospective StudiesABSTRACT
INTRODUCTION: Radial forearm flap donor side defects can be treated by vacuum therapy and conventional wound dressing. The aim of this study was to compare different wound management considering wound healing, including risk factors and cost effectiveness. MATERIALS AND METHODS: Retrospective study including patients treated with radial flaps in the Department of Head and Neck Surgery in Erlangen from January 2005 to August 2013. Wound healing was assessed regarding complications considering several risk factors and comorbidities. Data were analyzed comparing conventional dressing and vacuum therapy, including a calculation of costs. RESULTS: The study included 138 patients (n = 55 conventional dressing; n = 83 vacuum dressing). The incidence of wound complications in the vacuum group was 50.6 and 32.7% in the conventional dressing group (p = .058). The presence of risk factors and comorbidities did not have significant impact on the occurrence of complications. Costs for vacuum therapy turned out to be at least five times higher. CONCLUSION: In view of the current state of research, these results show that using vacuum dressings has no significant benefit in the wound management of forearm donor side defect covered with full-thickness skin grafts. If the costs and economical aspects are also considered, conventional wound dressing may be preferred.
Subject(s)
Negative-Pressure Wound Therapy , Surgical Flaps , Adult , Aged , Aged, 80 and over , Female , Forearm , Humans , Male , Middle Aged , Retrospective StudiesABSTRACT
BACKGROUND/AIM: Total glossectomy without laryngectomy for large tongue carcinomas still remains controversial, as these defects may go along with dramatic effects on respiration, speech and swallowing. As a consequence, these procedures significantly impact quality of life. Therefore, larger case series are rare. Nevertheless, with the development of free tissue transfer, functional reconstruction has become increasingly popular and encouraging results are reported in the literature. PATIENTS AND METHODS: We present our experience with complete glossectomy without laryngectomy and free flap reconstruction. A retrospective monocenter cohort-study was conducted. Reconstructive principles, which in our view lead to optimal functional results, are presented. Oncologic, as well as functional results are reported. Functional results were assessed in terms of swallowing ability, decannulation and intelligible speech. RESULTS: A total of 14 patients met the inclusion criteria. All patients were reconstructed using an anterolateral thigh flap. Complications occurred in 4 patients, 3 of which developed fistula formation. Oral feeding without the need for a gastrostomy tube was resumed in 11 patients (78.6%), 12 patients (85.7%) were able to be permanently decannulated and speech was at least acceptable in 12 patients (85.7%). The three-year survival was 57.1%. CONCLUSION: Following meticulous reconstructive principles, as well as a proper patient selection, total glossectomy without laryngectomy is a feasible treatment option for advanced cancer of the tongue.
Subject(s)
Glossectomy/methods , Neoplasms, Squamous Cell/surgery , Plastic Surgery Procedures/methods , Tongue Neoplasms/surgery , Adult , Aged , Disease-Free Survival , Female , Humans , Laryngectomy , Male , Middle Aged , Neoplasm Staging , Neoplasms, Squamous Cell/pathology , Postoperative Complications/pathology , Tongue Neoplasms/pathologyABSTRACT
The "Comprehensive ICF Core Set for Head and Neck Cancer" (ICF-HNC) is an application of the "International Classification of Functioning, Disability and Health" (ICF), representing the characteristic spectrum of issues in patients with head and neck cancer (HNC). Our primary aim was to evaluate which categories of the ICF-HNC are dealt with by speech and language therapists (SLTs) in Germany, Austria, and Switzerland. The secondary aim was to identify outcome measures used by SLTs to measure the categories of the ICF-HNC in clinical practice. SLTs experienced in the treatment of HNC patients evaluated the categories of the ICF-HNC in a three-round Delphi survey. They were asked whether the listed categories represented issues treated by SLTs in HNC patients, and what outcome measures were used to assess them. Altogether, 31 SLTs completed the survey. 47 of 108 previously selected categories of the ICF-HNC achieved the cut-off value. Out of these, 40.4% were derived from the component "Body Functions", 36.2% from "Body Structures", 12.8% from "Environmental Factors", and 10.6% from "Activities and Participation". Altogether, 82 of the mentioned outcome measures were considered as reasonable from the perspective of SLTs. Of these, only 37 achieved more than 50% approval. This study emphasises the importance of "Body Structures" and "Body Functions" for SLTs in Germany and Switzerland in treating patients with HNC. Moreover, the results highlighted the need to agree on evidence-based outcome measures in speech and language therapy.
Subject(s)
Disabled Persons/classification , Head and Neck Neoplasms/rehabilitation , Speech Disorders/classification , Speech-Language Pathology , Adult , Austria , Consensus , Delphi Technique , Disability Evaluation , Female , Germany , Humans , Language , Male , Middle Aged , Speech Disorders/etiology , Speech Disorders/rehabilitation , Speech Therapy , Surveys and Questionnaires , SwitzerlandABSTRACT
The assessment and evaluation of functioning and quality of life after tumor treatment in head and neck cancer (HNC) are considered as essential aspects of clinical routine and studies. A huge number of instruments are available that have been designed to evaluate functioning and quality of life after HNC treatment. The diversity of these instruments in terms of content, response options and administration hinders the comparability of available studies and the performance of meta-analyses. The objective of this paper is to inform about the development of a screening tool for the standardized assessment and evaluation of functioning based on the International Classification of Functioning, Disability and Health (ICF) Core Set for HNC. We followed a multi-step approach including (1) preparatory studies to identify and preselect suitable instruments for the assessment of functioning, (2) a decision-making process to agree on an ICF-based clinical guideline including instruments assessing functioning and (3) the development of a computer-based standardized screening tool to assess and evaluate functioning based on this guideline in clinical routine. Twenty-one experts participated in a consensus meeting and decided on instruments to be included in an ICF-based clinical guideline and screening tool for the assessment and evaluation of functioning in HNC patients in cancer treatment. The chosen instruments cover all aspects of the ICF Core Set for HNC addressing therapy control, pain, food intake/swallowing, voice/speech/breathing, other somatic complaints and psychosocial aspects. The screening tool contains patient-reported outcome measures and a clinician's checklist. It has to be further tested in clinical practice.
Subject(s)
Consensus , Disability Evaluation , Disabled Persons/rehabilitation , Head and Neck Neoplasms/rehabilitation , Mass Screening/methods , Practice Guidelines as Topic , Humans , Quality of LifeSubject(s)
Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/virology , Oropharyngeal Neoplasms/diagnosis , Oropharyngeal Neoplasms/virology , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Adult , Carcinoma, Squamous Cell/epidemiology , Cross-Sectional Studies , Early Diagnosis , Female , Human papillomavirus 16 , Humans , Incidence , Male , Middle Aged , Oropharyngeal Neoplasms/epidemiology , Papillomavirus Infections/epidemiology , Prognosis , Sexual Behavior , Smoking/adverse effectsABSTRACT
PURPOSE: To assess risk factors of patients with oropharyngeal squamous cell carcinoma in the Munich area of Southern Germany in relation to human papillomavirus (HPV) association of the tumors. To demonstrate differences in tumor characteristics and their impact on adjuvant treatment. METHODS: Between November 2010 and July 2013, patients were prospectively interviewed for risk factors before they underwent surgical resection of their tumors. HPV association was evaluated by p16 immunohistochemistry; tumor characteristics and type of adjuvant treatment were recorded. Follow-up data were collected after a median follow-up of 12.1 month. RESULTS: In contrast to many recent studies, we could not detect any difference in overall age and age at sexual debut between p16-positive and p16-negative patients. P16-negative patients are characterized by a more intensive tobacco and alcohol use, a more abusive way of consumption, less nonoral and less oral sex partners. P16-positive patients had a significantly higher risk of lymph node metastases, but nevertheless a significant lower risk to recur or to die. No difference in the incidence of synchronous second primary tumors was seen. P16-positive patients generally received a more aggressive adjuvant treatment because of more frequently involved lymph nodes. CONCLUSION: Lifestyle risk factors such as smoking, drinking, and sexual habits were independent from age, but showed marked differences between the p16-positive and p16-negative group. Since p16-positive patients were treated more aggressively, it is not possible to distinguish whether the better outcome of HPV-positive patients is a result of less aggressive cancers or more aggressive treatment. With regard to the ongoing debate about treatment deintensification, we should keep in mind that the survival of HPV-positive cancer patients is not 100 %.
Subject(s)
Carcinoma, Squamous Cell/surgery , Oropharyngeal Neoplasms/surgery , Adult , Aged , Aged, 80 and over , Alcohol Drinking/adverse effects , Biomarkers, Tumor/analysis , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/virology , Cyclin-Dependent Kinase Inhibitor p16/metabolism , DNA, Viral/genetics , Female , Follow-Up Studies , Germany , Humans , Male , Middle Aged , Neoplasm Grading , Neoplasm Invasiveness , Neoplasm Staging , Oropharyngeal Neoplasms/pathology , Oropharyngeal Neoplasms/virology , Papillomaviridae/genetics , Papillomavirus Infections/pathology , Papillomavirus Infections/surgery , Papillomavirus Infections/virology , Prognosis , Prospective Studies , Risk Factors , Smoking/adverse effectsABSTRACT
UNLABELLED: The head and neck region is one of the most important locations predisposed for tobacco-associated cancer. Chemoprevention might offer a chance to decrease the risk for this type of disease. MATERIALS AND METHODS: Mini-organ cultures (MOC) of macroscopically-healthy pharyngeal tissues from 20 patients with oropharyngeal squamous cell carcinoma (SCC) and from 20 controls were employed in the study. MOC were firstly incubated with Celecoxib, and DNA damage was induced by incubation with Benz[a]pyren-7,8-diol-9,10-epoxid (BPDE), a major representative of tobacco-associated carcinogens. DNA damage was evaluated with the alkaline single-cell microgel electrophoresis (Comet assay). Furthermore, fragmentation of the cyclin D1 gene, a gene of special importance in head and neck carcinogenesis was examined by the Comet-FISH assay. Finally, the chemoprotective potential of Celecoxib was analyzed after incubation with MOC. RESULTS: As expected, BPDE caused significant DNA fragmentation in tumor compared to negative control tissues. No enhanced damage was observed in the cyclin D1 gene. DNA fragmentation was significantly reduced when MOC were incubated with Celecoxib in the tumor group. Surprisingly, these effects were also observed in the group without cancer of the oropharynx, although COX-2 is not expressed in macroscopically-healthy mucosa. CONCLUSION: Celecoxib showed considerable chemoprotective effeciency against BPDE in both groups and this effect seems to be independent of COX-2 expression. No evidence for higher mutagen sensitivity in the Cyclin D1 gene was observed.
Subject(s)
Carcinoma, Squamous Cell/genetics , Cyclin D1/metabolism , Head and Neck Neoplasms/genetics , Oropharyngeal Neoplasms/genetics , Pyrazoles/administration & dosage , Sulfonamides/administration & dosage , Adult , Aged , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/pathology , Celecoxib , Chemoprevention , Cyclooxygenase 2/biosynthesis , DNA Damage/drug effects , DNA Fragmentation/drug effects , Female , Gene Expression Regulation, Neoplastic/drug effects , Head and Neck Neoplasms/drug therapy , Head and Neck Neoplasms/pathology , Humans , Male , Middle Aged , Mutagens/toxicity , Organ Culture Techniques , Oropharyngeal Neoplasms/drug therapy , Oropharyngeal Neoplasms/pathology , Tobacco Use Disorder/drug therapy , Tobacco Use Disorder/genetics , Tobacco Use Disorder/pathologyABSTRACT
The goals of the present publication are (a) to identify standardised methods and tools applied in clinical trials and cohort studies to assess key functional aspects in HNC, using the Brief ICF Core Set for head and neck cancer (ICF-HNC) as a reference and (b) to propose a set of sound standardised methods and tools suitable to assess functional problems in HNC. This work contributed to the development of practical ICF-HNC based guidelines targeting the standardised measurement of functional outcomes in HNC follow-up and clinical research in Germany. A systematic review of randomised and clinical controlled trials, and observational studies in HNC were carried out to identify standardised methods and tools. Suitable methods and tools were then selected based on pre-defined criteria. 210 assessment methods and tools were identified in 136 studies: 146 patients reported outcomes (PRO) and 64 tools rated by health professionals. Altogether 59 tools were considered suitable to be included in guidelines: four side effects classifications, two performance status scales, 31 PROs, 10 assessment criteria for clinical examinations, seven assessment methods and tools for the evaluation of technical, equipment-based procedures and five technical, equipment-based procedures. It was possible to identify and select sound and standardised assessment methods and tools for almost all functioning areas defined in the ICF-HNC. Since no tool sufficiently covers support provision by immediate family and by health professionals as well as economic self-sufficiency, we recommend a corresponding update of existing tools.
Subject(s)
Activities of Daily Living , Head and Neck Neoplasms/physiopathology , Head and Neck Neoplasms/psychology , Patient Outcome Assessment , Recovery of Function , Clinical Trials as Topic , Cohort Studies , Disability Evaluation , Germany , Head and Neck Neoplasms/therapy , Humans , Quality of Life , Surveys and QuestionnairesABSTRACT
Google Trends is a valuable new additional tool for medical research, mainly for epidemiological and economical issues, which is fast and inexpensive to use and may be especially helpful when analyzing patient collectives who go to general practitioners or specialists in private practice rather than public or university clinics, where data for statistics are most often collected from.
Subject(s)
Information Seeking Behavior , Otolaryngology , Web Browser , Consumer Health Information , Internet , Statistics as TopicABSTRACT
The International Classification of Functioning, Disability and Health Core Set for Head and Neck Cancer (ICF-HNC) covers the typical spectrum of problems in functioning experienced by patients with head and neck cancer (HNC). The major goal of the present work was to evaluate patients' priorities using the brief ICF-HNC as a starting point. A priorities assessment checklist consisting of 15 statements was created based on the 14 validated categories of the brief ICF-HNC. In a cross-sectional study, patients were requested to select up to 5 items that were especially important to them. The checklist was sent by mail to 465 patients at different time points of cancer follow-up and handed out to 56 patients with recent HNC diagnosis. Altogether 300 (64.51 %) patients returned the checklist. The top priority of our sample was "I want to survive the cancer", followed by "I want that all the expenses for cancer treatment, cancer care and any additional follow-up treatments be covered by my health insurance or by the welfare system", "I want to be able to continue performing all daily life tasks well", "I want to have trusting relationships with my doctors, nurses and therapists" and "I want to be able to speak clearly". Although survival was, as expected, the top priority for patients enrolled in the study, we show that the weight given to survival and further symptoms or daily life activities meaningfully changes when the biopsychosocial perspective proposed in the ICF is adopted.
Subject(s)
Head and Neck Neoplasms/psychology , Health Priorities , Quality of Life , Adult , Cross-Sectional Studies , Disability Evaluation , Female , Head and Neck Neoplasms/pathology , Head and Neck Neoplasms/therapy , Humans , Male , Neoplasm Staging , Outcome Assessment, Health Care , Recovery of Function , Surveys and QuestionnairesABSTRACT
BACKGROUND: Since the middle of the 1990s vacuum-assisted closure (VAC) has been used in many areas of surgery to manage complex wounds and impaired wound healing. Until recently, little attention has been paid to this treatment modality in the field of head and neck surgery. The evaluation of its efficacy in wound healing disorders of the head and neck was the aim of this study. MATERIAL AND METHODS: Patients with complex wounds and impaired healing treated with VAC therapy between 2008 and 2011 were included into the study. VAC dressings were changed every 3 days and improvements in wound healing were documented. RESULTS: 23 patients were treated with VAC therapy, in 18 cases (78%) closure of the defect could be reached without any further surgical procedure. 5 patients needed subsequent regional flap reconstruction to close the remaining defect. All of these patients had undergone salvage surgery in a previously irradiated neck before. CONCLUSION: Vacuum assisted closure is an effective treatment in the management of wound healing disorders and complex wounds in the head and neck. It offers a useful, non-invasive modality to close even large defects in the area. Previous irradiation seems to have a significant negative influence on the outcome of the therapy, but more data are required to assess these effects.
Subject(s)
Head and Neck Neoplasms/surgery , Negative-Pressure Wound Therapy/methods , Plastic Surgery Procedures/adverse effects , Surgical Wound Infection/therapy , Wound Healing , Adult , Aged , Female , Follow-Up Studies , Humans , Male , Middle Aged , Retrospective Studies , Surgical Flaps , Treatment OutcomeABSTRACT
The study of tumourigenesis commonly involves the use of established cell lines or single cell suspensions of primary tumours. Standard methods for the generation of short-term tumour cell cultures include the disintegration of tissue based on enzymatic and mechanical stress. Here, we describe a simple and rapid method for the preparation of single cells from primary carcinomas, which is independent of enzymatic treatment and feeder cells. Tumour biopsies are processed to 1 mm(3) cubes termed explants, which are cultured 1-3 days on agarose-coated well plates in specified medium. Through incisions generated in the explants, single cells are retrieved and collected from the culture supernatant and can be used for further analysis including in vitro and in vivo studies. Collected cells retain tumour-forming capacity in xenotransplantation assays, mimic the phenotype of the primary tumour, and facilitate the generation of cell lines.