Background Conducting research on the molecular biology, immunology, and physiology of brain tumors (BTs) and primary brain tissues requires the use of viably dissociated single cells. Inadequate methods for tissue dissociation generate considerable loss in the quantity of single cells produced and in the produced cells’ viability. Improper dissociation may also demote the quality of data attained in functional and molecular assays due to the presence of large quantities cellular debris containing immune-activatory danger associated molecular patterns, and due to the increased quantities of degraded proteins and RNA.ResultsOver 40 resected BTs and non-tumorous brain tissue samples were dissociated into single cells by mechanical dissociation or by mechanical and enzymatic dissociation. The quality of dissociation was compared for all frequently used dissociation enzymes (collagenase, DNase, hyaluronidase, papain, dispase) and for neutral protease (NP) from Clostridium histolyticum. Single-cell-dissociated cell mixtures were evaluated for cellular viability and for the cell-mixture dissociation quality. Dissociation quality was graded by the quantity of subcellular debris, non-dissociated cell clumps, and DNA released from dead cells. Of all enzymes or enzyme combinations examined, NP (an enzyme previously not evaluated on brain tissues) produced dissociated cell mixtures with the highest mean cellular viability: 93 % in gliomas, 85 % in brain metastases, and 89 % in non-tumorous brain tissue. NP also produced cell mixtures with significantly less cellular debris than other enzymes tested. Dissociation using NP was non-aggressive over time—no changes in cell viability or dissociation quality were found when comparing 2-h dissociation at 37 °C to overnight dissociation at ambient temperature.ConclusionsThe use of NP allows for the most effective dissociation of viable single cells from human BTs or brain tissue. Its non-aggressive dissociative capacity may enable ambient-temperature shipping of tumor pieces in multi-center clinical trials, meanwhile being dissociated. As clinical grade NP is commercially available it can be easily integrated into cell-therapy clinical trials in neuro-oncology. The high quality viable cells produced may enable investigators to conduct more consistent research by avoiding the experimental artifacts associated with the presence dead cells or cellular debris.Electronic supplementary materialThe online version of this article (doi:10.1186/s12868-016-0262-y) contains supplementary material, which is available to authorized users.

Object Orbital tumors can be divided schematically into primary lesions, originating from the orbit itself, and secondary lesions, extending to the orbit from neighboring structures. These tumors are variable in their biological nature and in their location. The authors evaluate 41 cases of benign and malignant tumors involving the orbit and discuss the surgical challenge, which involves tumor removal, preserving visual function and cosmetic reconstruction. Methods The authors performed a retrospective analysis of a series of all cases involving patients who underwent surgery for treatment of orbital tumors in their hospital between December 2003 and December 2006. Data were collected from the patients' files in the hospital's outpatient clinic, operative notes, and pre- and postoperative imaging studies. Results The authors identified 41 patients who met the inclusion criteria (age range 14–82 years, mean 42.2 years, standard deviation 22.4 years). The most common presenting symptoms were proptosis and/or partial ptosis (11 cases [27%]) and headache (7 cases [17%]). In 5 (12%) cases, the tumors were primary intraorbital lesions, and in 34 cases (83%) they were secondary. Two patients had metastases to the orbit. The most common lesion types were meningioma (10 cases [24%]) and osteoma (7 cases [17%]). In 24 cases a midline approach through a frontal craniotomy or a subcranial approach was used; in 13 a lateral approach was used; and in 4 a biopsy procedure was performed. Maxillectomy through a Weber–Ferguson approach or a facial degloving approach was added in 5 cases to complete tumor removal. Duration of follow-up was 1–38 months (mean 20 months). None of the patients died as a result of the procedure, and there were relatively few complications. Excluding the patients who underwent orbital exenteration, none of the patients had visual deterioration following surgery, and most had no change in their visual condition. Two patients had temporary diplopia, 1 had a cerebrospinal fluid leak, and 1 had enophthalmos following removal of an orbital osteosarcoma. Conclusions Orbital tumors can be treated safely using transcranial approaches in many cases. Preoperative imaging can accurately define the compartments involved and the surgical approach needed for tumor removal. A multidisciplinary team of surgeons facilitates optimal tumor removal and skull base sealing as well as good cosmetic results.

Tuberculum and diaphragma sella meningioma can be safely resected using the subfrontal approach with preservation and even improvement of visual function after surgery. Early surgery with better pre-operation visual function and smaller tumour size were associated with a better outcome.

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BACKGROUND AND PURPOSE:The artery of Bernasconi and Cassinari is an important infraclinoid branch of the internal cerebral artery. It is of neuroendovascular relevance in view of its supply to complex lesions such as meningiomas and arteriovenous malformations in the tentorial and falcotentorial regions. The present microanatomic study attempts a morphometric elucidation of this slender but important branch of the meningohypophyseal trunk.