# US 10,481,151 B2 · Isolation of pulmonary arterial endothelial cells from patients with pulmonary vascular disease and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,481,151 B2 - Document ID: US10481151B2 - Status: Granted - Kind: B2 - Application number: 15/806,751 - Filed: 2017-11-08 - Published: 2018-03-08 - Granted: 2019-11-19 - Earliest claimed: 2012-09-10 - Assignees: Allegheny-Singer Research Institute - Applicants: Allegheny-Singer Research Institute - Inventors: Michael J. Passineau; Srinivas Murali; Raymond L. Benza; Jonathan B. Pollett - Web page: https://patentgrep.com/patent/US10481151B2 ## CPC - G01N 33/5091: {for testing the pathological state of an organism} - A61B 10/02: Instruments for taking cell samples or for biopsy {(A61B10/0038 and A61B10/0045 take precedence; needle locating or guiding means A61B17/3403; samplers for enzymology or microbiology C12M1/26; sampling or preparing biological specimens G01N33/48)} - A61M 25/10: Balloon catheters ({A61M25/0125 takes precedence; embolectomy A61B17/22032; retractors A61B17/02;} inflatable balloons for placing stents or stent-grafts A61F2/958 {; stomach balloons for treatment of obesity A61F5/0003; oesophagal tubes A61J15/00}) - C12N 5/069: {Vascular Endothelial cells} - A61M 2202/097: endothelial cells - G01N 2570/00: Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes - G01N 2800/321: Arterial hypertension - G01N 2800/60: Complex ways of combining multiple protein biomarkers for diagnosis - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present disclosure provides methods of obtaining, recovering and culturing pulmonary arterial endothelial cells from an individual patient subjected to a right heart catheterization in regard to diagnosis of pulmonary hypertension. These isolated pulmonary arterial endothelial cells from an individual patient may be utilized to generate cellular phenotyping profile for the cell population, allowing generation of a personalized pharmacotherapy regime to assist the clinician in treatment and prognosis of the disorder for an individual patient. ## Claims ### Claim 1 A method of culturing a population of pulmonary arterial endothelial cells from a patient, comprising: introducing an inflatable balloon catheter into a vein of the patient and advancing the catheter through the right heart of the patient and into a branch of the pulmonary artery of the patient; inflating the balloon catheter to a pressure consistent with that of a pulmonary artery catheter; removing the balloon catheter from the patient; collecting a total cell population that adheres to the balloon catheter after removal from the patient; purifying the population of pulmonary endothelial cells away from the total cell population; and expanding the population of pulmonary endothelial cells in a growth medium. ### Claim 2 (depends on 1) The method of claim 1, wherein the patient shows symptoms of or is diagnosed with pulmonary hypertension. ### Claim 3 (depends on 2) The method of claim 2, wherein the patient shows symptoms of or is diagnosed with a form of pulmonary hypertension selected from the group consisting of pulmonary arterial hypertension, left-sided atrial heart disease, left-sided ventricular heart disease, pulmonary hypertension associated with lung diseases and/or hypoxemia, pulmonary hypertension due to chronic thrombotic and/or embolic disease, sarcoidosis, histiocytosis X, lymphangiomatosis, adenopathy associated with compression of pulmonary vessels, tumor presence associated with compression of pulmonary vessels, and fibrosing mediastinitis associated with compression of pulmonary vessels. ### Claim 4 (depends on 1) The method of claim 1, wherein the pulmonary arterial endothelial cells express CD31, CD146, CD105 and/or CD133, and/or are CD45 negative. ### Claim 5 (depends on 1) The method of claim 1, further comprising passaging of the pulmonary arterial endothelial cells in culture. ### Claim 6 (depends on 1) The method of claim 1, wherein performing the cell purification process comprises using flow cytometry, magnetic-based cell separation, or fluorescence activated cell sorting to separate the pulmonary arterial endothelial cells away from the total cell population. ### Claim 7 (depends on 1) The method of claim 1, further comprising performing one or more in vitro cell-based screens or assays on the pulmonary arterial endothelial cells to measure one or a combination of biomarkers associated with a pulmonary hypertension disorder. ### Claim 8 (depends on 7) The method of claim 7, further comprising: based on the one or more in vitro cell-based screens or assays, selecting and providing to the patient a personalized pharmacotherapy regime for the patient concerning the treatment of a pulmonary hypertension disorder. ### Claim 9 (depends on 8) The method of claim 8, wherein the in vitro cell-based screen or assay comprises measurement of Bcl-2 or BMPR-II expression. ### Claim 10 (depends on 1) The method of claim 1, further comprising performing one or more in vitro cell-based screens or assays to measure: cellular phenotypic characteristics indicative of a stage of pulmonary hypertension in the population of pulmonary arterial endothelial cells; or cellular phenotypic characteristics indicative of detectable transformation of at least a threshold percentage of the population of pulmonary arterial endothelial cells from a normal cellular apoptotic state to a hyper-apoptotic state or an anti-apoptotic state. ### Claim 11 (depends on 10) The method of claim 10, wherein: a determination that at least a threshold percentage of the population of pulmonary arterial endothelial cells are in a hyper-apoptotic state indicates that the patient exhibits an early stage of pulmonary vascular disease; and a determination that at least a threshold percentage of the population of pulmonary arterial endothelial cells are in an anti-apoptotic state indicates that the patient exhibits a late stage of pulmonary vascular disease. ### Claim 12 A method of culturing a population of pulmonary arterial endothelial cells from a patient, comprising: introducing an inflatable balloon catheter into a vein of a patient who shows symptoms of or is diagnosed with pulmonary hypertension and advancing the catheter through the right heart of the patient and into a branch of the pulmonary artery of the patient; inflating the balloon catheter to a pressure consistent with that of a pulmonary artery catheter; removing the balloon catheter from the patient; collecting a total cell population that adheres to the balloon catheter after removal from the patient; using flow cytometry, magnetic-based cell separation, or fluorescence activated cell sorting to separate the pulmonary arterial endothelial cells away from the total cell population; and expanding the population of pulmonary endothelial cells in a growth medium. ### Claim 13 (depends on 12) The method of claim 12, wherein the patient shows symptoms of or is diagnosed with a form of pulmonary hypertension selected from the group consisting of pulmonary arterial hypertension, left-sided atrial heart disease, left-sided ventricular heart disease, pulmonary hypertension associated with lung diseases and/or hypoxemia, pulmonary hypertension due to chronic thrombotic and/or embolic disease, sarcoidosis, histiocytosis X, lymphangiomatosis, adenopathy associated with compression of pulmonary vessels, tumor presence associated with compression of pulmonary vessels, and fibrosing mediastinitis associated with compression of pulmonary vessels. ### Claim 14 (depends on 12) The method of claim 12, wherein the pulmonary arterial endothelial cells express CD31, CD146, CD105 and/or CD133, and/or are CD45 negative. ### Claim 15 (depends on 12) The method of claim 12, further comprising passaging of the pulmonary arterial endothelial cells in culture. ### Claim 16 (depends on 12) The method of claim 12, further comprising performing one or more in vitro cell-based screens or assays on the pulmonary arterial endothelial cells to measure one or a combination of biomarkers associated with a pulmonary hypertension disorder. ### Claim 17 (depends on 16) The method of claim 16, further comprising: based on the one or more in vitro cell-based screens or assays, selecting and providing to the patient a personalized pharmacotherapy regime for the patient concerning the treatment of a pulmonary hypertension disorder. ### Claim 18 (depends on 17) The method of claim 17, wherein the in vitro cell-based screen or assay comprises measurement of Bcl-2 or BMPR-II expression. ### Claim 19 (depends on 12) The method of claim 12, further comprising performing one or more in vitro cell-based screens or assays to measure: cellular phenotypic characteristics indicative of a stage of pulmonary hypertension in the population of pulmonary arterial endothelial cells; or cellular phenotypic characteristics indicative of detectable transformation of at least a threshold percentage of the population of pulmonary arterial endothelial cells from a normal cellular apoptotic state to a hyper-apoptotic state or an anti-apoptotic state. ### Claim 20 (depends on 12) The method of claim 12, further comprising performing one or more in vitro cell-based screens or assays to determine whether at least a threshold percentage of the population of pulmonary arterial endothelial cells has transformed from a normal cellular apoptotic state to a hyper-apoptotic state or an anti-apoptotic state, wherein: a determination that at least a threshold percentage of the population of pulmonary arterial endothelial cells are in a hyper-apoptotic state indicates that the patient exhibits an early stage of pulmonary vascular disease; and a determination that at least a threshold percentage of the population of pulmonary arterial endothelial cells are in an anti-apoptotic state indicates that the patient exhibits a late stage of pulmonary vascular disease.