Harvest of pulmonary artery endothelial cells from patients undergoing right heart catheterization
| Autor: | Jonathan B. Pollett, Srinivas Murali, Kelly J. Shields, Michael J. Passineau, Raymond L. Benza |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Pulmonary and Respiratory Medicine
Cardiac Catheterization Pathology medicine.medical_specialty Hypertension Pulmonary medicine.medical_treatment Context (language use) Pulmonary Artery Balloon medicine.artery medicine Humans Familial Primary Pulmonary Hypertension Endothelial dysfunction Saline Transplantation business.industry Endothelial Cells medicine.disease Endothelial stem cell Catheter medicine.anatomical_structure Pulmonary artery Surgery Cardiology and Cardiovascular Medicine business Artery |
| Zdroj: | The Journal of Heart and Lung Transplantation. 32:746-749 |
| ISSN: | 1053-2498 |
| Popis: | Endothelial dysfunction is a hallmark of pulmonaryarterial hypertension (PAH). Although the exact roleendothelial dysfunction plays in the initiation or progressionof PAH is not yet clear, impaired apoptosis and aberrantsignaling in endothelial cells is observed in most forms ofPAH. A variety of approaches have been utilized to studyendothelial cell biology in the context of PAH. Most studieshave focused on in situ visualization or purification ofpulmonary artery endothelial cells (PAECs) from explantedlungs of PAH patients, using untransplanted human donorlungs as controls. Although very useful for comparingendothelial cell biology in PAH versus normal pulmonaryvasculature, this approach is unable to provide informationon dynamic changes in PAEC biology over time within aliving patient.The definitive diagnosis of PAH requires measurement ofmean pulmonary artery (PA) pressure via right heartcatheterization (RHC), and thus RHC serves as a “goldstandard” for diagnosis and as a gateway for the manage-ment of PAH patients. We hypothesized that PAECs mightbe dislodged from the PA vessel wall by the balloon tip ofthe flow-directed pulmonary artery (Swan–Ganz) catheterduring RHC and would remain adherent to the balloon whenit is collapsed and the catheter extracted. Herein we reportour successful efforts to isolate, characterize and expandPAECs recovered from Swan–Ganz catheter balloons afterroutine RHC for the diagnosis and management of PAH.After obtaining institutional review board approval, 24patients undergoing RHC consented to participate in ourstudy. After the RHC procedure, the last 2 inches of theSwan–Ganz catheter, including the inflatable balloon, wascut and placed in a tube containing endothelial cell media(Cell Applications, Inc., San Diego, CA) and placed on iceuntil the samples were further processed. In the laboratory,catheter tips were washed vigorously 5 with phosphate-buffered saline (PBS)/trypsin and centrifuged at 1,500 rpmfor 10 minutes. The resulting pellets were re-suspended inammonium–chloride–potassium (ACK)/PBS solution tolyse the erythrocytes. These suspensions were centrifugedat 2,500 rpm for 10 minutes to pellet the cells. An aliquot ofthis final cell harvest was analyzed by fluorescence-activated cell sorting (FACS) with antibodies for endothelialantigens and the balance transferred to primary culture.In these studies, we relied on the classic marker profile forendothelial cells as reported in 2001 by Mancuso et al |
| Databáze: | OpenAIRE |
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