microvascular-macrovascular-endothelial-cell-isolation-purification

Microvascular and Macrovascular Endothelial Cell Isolation and Purification from Lung-Derived Samples

The availability of cells isolated from healthy and diseased tissues and organs represents a key element for personalized medicine approaches. Although ...

Endothelial cell (EC) dysfunction has been reported in cystic fibrosis (CF) patients. Thus, the availability of CF EC is paramount to uncover mechanisms of endothelial dysfunction in CF. Using collagenase digestion, we isolated cells from small fragments of pulmonary artery dissected from non-CF lobes or explanted CF lungs. These cells were a heterogeneous mixture, containing variable percentages of EC. To obtain virtually pure pulmonary artery endothelial cells (PAEC), we developed an easy, inexpensive, and reliable method, based on the differential adhesion time of pulmonary artery cells collected after collagenase digestion. With this method, we obtained up to 95% pure non-CF and CF-PAEC. Moreover, we also succeed at immortalizing both PAEC and CF-PAEC, which remained viable and with unchanged phenotype and proliferation rate over the 30th passage. These cells recapitulated cystic fibrosis transmembrane conductance regulator expression and functions of the parental cells. Thus, we isolated for the first time endothelial cells from CF patients, providing a valuable tool to define the emerging role of EC in CF lung and vascular disease.

Establishment and long-term culture of human cystic fibrosis endothelial cells.

The transcriptional profiling of cancer and normal tissues harboring cancer can be a clinical and discovery tool, especially for the study of rare tumors. Invasive mucinous adenocarcinoma (IMA) is a rare lung cancer histotype, which mostly affects the elderly and commonly has a poor prognosis. We investigated the exceptional case of a teenager, exposed to passive smoke and chemical carcinogens, who developed a multifocal IMA with bilateral involvement. The malignancy was asymptomatic and was diagnosed occasionally during hospitalization for acute abdominal pain due to adnexitis. The young patient underwent video-assisted thoracoscopic surgery and lung samples were analysed by RNA-Sequencing. The transcriptome of patient's normal and neoplastic lung tissues was compared with matched healthy controls and IMA signature cases, using Gene Set Enrichment Analyses, Gene Ontology and Genotype Tissue Expression database. Compared to healthy controls, the patient's lung tissue lacked the expression of lymphocyte and humoral-mediated immune response genes, whereas genes driving the response to stimulus, chemical and organic substances, primarily, , , , ,  and  were overexpressed. Genes driving extracellular structure organization, cell adhesion, cell movement, metabolic and apoptotic processes were down-modulated in patient's lung tissue. When compared to IMA signature cases, the patient's IMA revealed a prevalent expression of genes regulating the response to stimulus, myeloid and neutrophil activation and immune system processes, primarily  and , whereas stemness genes and proto-oncogenes, such as , ,  and  were downmodulated. These transcriptional signature associated with a favorable clinical course, since the patient was healthy five years after initial diagnosis. The transcriptome of the normal tissues bearing tumor provides meaningful information on the gene pathways driving tumor histogenesis, with a prospective impact on early diagnosis. Unlike the tumor histotype-related transcriptional signature, the individual patient's signature enables tailored treatment and accurate prognosis.

Clinical impact of the lung tissue transcriptome in a teenager with multifocal invasive mucinous adenocarcinoma-a case report.

[This corrects the article DOI: 10.21037/tlcr-20-177.].

Erratum to clinical impact of the lung tissue transcriptome in a teenager with multifocal invasive mucinous adenocarcinoma-a case report.

Although laparoscopic cholecystectomy (LC) is the gold standard for symptomatic gallbladder disease, a single-incision approach may be a new challenge in order to achieve minimization of surgical trauma. Single-site robotic cholecystectomy (SSRC) is able to offset the ergonomic limitation of laparoscopic single-site cholecystectomy and improves cosmesis.

Outcome after single-site robotic cholecystectomy: An initial single center's experience.

Gallbladder agenesis (GA) is a rare embryological anomaly that presents acute cholecystitis like-symptoms. It is often an incidental finding diagnosed during surgery. We reported a case of GA in a patient who presented with dyspepsia and acute right upper abdomen pain with ultrasonographic signs of acute lithiasic cholecystitis. The preoperative assessment, according to first-level exams, is oriented to the diagnosis of acute lithiasic cholecystitis with atrophy and sclerosis. During laparoscopy, the proximal transverse colon was found strictly adherent to gallbladder fossa. The gallbladder was found to be absent. The surgical procedure consisted of lysis of multiple colo-hepatic adhesions. The diagnosis of congenital GA was made laparoscopically. The postoperative radiological images, based on CT and MR examinations, documented the diagnosis of GA with a biliary duct anatomical variant. The recovery was uneventful and the patient remained symptom-free for more than four years. GA is a clinical challenge that still poses diagnostic and therapeutic dilemmas. Although no diagnostic and therapeutic algorithm is accepted worldwide, due to heterogeneity of clinical scenarios and the variability in hospital facilities, surgeons have to be familiar with this rare entity, and conversion in laparotomy or unnecessary operative procedures should be avoided in the same operative setting.

Article	Year
Establishment and long-term culture of human cystic fibrosis endothelial cells. Laboratory investigation; a journal of technical methods and pathology\| PubMed ID: 28759010	2017
Clinical impact of the lung tissue transcriptome in a teenager with multifocal invasive mucinous adenocarcinoma-a case report. Translational lung cancer research\| PubMed ID: 32676340	2020
Erratum to clinical impact of the lung tissue transcriptome in a teenager with multifocal invasive mucinous adenocarcinoma-a case report. Translational lung cancer research\| PubMed ID: 33209650	2020
Outcome after single-site robotic cholecystectomy: An initial single center's experience. Asian journal of endoscopic surgery\| PubMed ID: 33264814	2021
A Case Report on Gallbladder Agenesis: Not a Novelty but Still a Laparoscopic Surprise. Cureus\| PubMed ID: 35047247	2021

Felice Mucilli

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