Highlights

• The endothelial cells of the pulmonary artery are a deficient model object. Our task was to work out the method of cell isolation from the Swan-Ganz catheter described in the literature. However, no cells were obtained using this technique. We have developed an alternative technique for isolating cells from blood obtained from the lumen of the pulmonary artery during right heart catheterization, and demonstrated the suitability of this technique for routine production of endothelial cell cultures from patients with pulmonary arterial hypertension.

Abstract

Aim. Obtaining cultures of pulmonary artery endothelial cells as a model object. The endothelium ensures the integrity of the vascular wall, and disorders of endothelial function play an important role in the development of various pathological processes. Obtaining endothelial cell cultures from fragments of various human vessels is a fairly well–developed and widely used method. However, it has limitations that relate to the availability of biological material for certain groups of patients. Thus, when studying the pathogenesis of pulmonary hypertension, fragments of the pulmonary artery are practically inaccessible to the researcher, since surgical intervention for this pathology is not a routine method of treatment.

Methods. The cells were obtained from the balloon of the Swan-Ganz catheter in accordance with the descriptions in the literature, alternative tissue dissociators and different adhesive media were additionally tested. The isolation of cells from the blood from the lumen of the pulmonary artery and from the peripheral (cubital) vein was carried out by dividing the blood into fractions and washing the cells of interest according to the protocols described in the literature.

Results. In this work, we tested the production of pulmonary artery endothelial cells in patients with pulmonary arterial hypertension by removing cells from the balloon of the Swan-Ganz catheter during right heart catheterization. The catheter removal method has shown its unsuitability for use in routine laboratory practice. As an alternative, we have developed a method for obtaining endothelial cells from blood taken from the lumen of the pulmonary artery during catheterization.

Conclusion. Isolation of endothelial cells from blood taken from the lumen of the pulmonary artery during right heart catheterization showed the suitability of this method for routine production of endothelial cell cultures from patients with pulmonary arterial hypertension.

1. Humbert, M., Sitbon, O., Guignabert, C., Savale, L., Boucly, A., Gallant-Dewavrin, M., McLaughlin, V., Hoeper, M. M., & Weatherald, J. (2023). Treatment of pulmonary arterial hypertension: Recent progress and a look to the future. The Lancet. Respiratory Medicine, 11(9), 804–819. https://doi.org/10.1016/S2213-2600(23)00264-3

2. Ruopp, N. F., & Cockrill, B. A. (2022). Diagnosis and Treatment of Pulmonary Arterial Hypertension: A Review. JAMA, 327(14), 1379–1391. https://doi.org/10.1001/jama.2022.4402

3. Evans, C. E., Cober, N. D., Dai, Z., Stewart, D. J., & Zhao, Y.-Y. (2021). Endothelial cells in the pathogenesis of pulmonary arterial hypertension. The European Respiratory Journal, 58(3), 2003957. https://doi.org/10.1183/13993003.03957-2020

4. Awad, K. S., Wang, S., Dougherty, E. J., Keshavarz, A., Demirkale, C. Y., Yu, Z. X., Miller, L., Elinoff, J. M., & Danner, R. L. (2024). BMPR2 Loss Activates AKT by Disrupting DLL4/NOTCH1 and PPARγ Signaling in Pulmonary Arterial Hypertension. International Journal of Molecular Sciences, 25(10), 5403. https://doi.org/10.3390/ijms25105403

5. Schupp, J. C., Adams, T. S., Cosme, C., Raredon, M. S. B., Yuan, Y., Omote, N., Poli, S., Chioccioli, M., Rose, K.-A., Manning, E. P., Sauler, M., DeIuliis, G., Ahangari, F., Neumark, N., Habermann, A. C., Gutierrez, A. J., Bui, L. T., Lafyatis, R., Pierce, R. W., … Kaminski, N. (2021). Integrated Single-Cell Atlas of Endothelial Cells of the Human Lung. Circulation, 144(4), 286–302. https://doi.org/10.1161/CIRCULATIONAHA.120.052318

6. Singh, N., Eickhoff, C., Garcia-Agundez, A., Bertone, P., Paudel, S. S., Tambe, D. T., Litzky, L. A., Cox-Flaherty, K., Klinger, J. R., Monaghan, S. F., Mullin, C. J., Pereira, M., Walsh, T., Whittenhall, M., Stevens, T., Harrington, E. O., & Ventetuolo, C. E. (2023). Transcriptional profiles of pulmonary artery endothelial cells in pulmonary hypertension. Scientific Reports, 13(1), 22534. https://doi.org/10.1038/s41598-023-48077-6

7. Zhou, Y., Tabib, T., Huang, M., Yuan, K., Kim, Y., Morse, C., Sembrat, J., Valenzi, E., & Lafyatis, R. (2024). Molecular Changes Implicate Angiogenesis and Arterial Remodeling in Systemic Sclerosis-Associated and Idiopathic Pulmonary Hypertension. Arteriosclerosis, Thrombosis, and Vascular Biology, 44(8), e210–e225. https://doi.org/10.1161/ATVBAHA.123.320005

8. Cober, N. D., VandenBroek, M. M., Ormiston, M. L., & Stewart, D. J. (2022). Evolving Concepts in Endothelial Pathobiology of Pulmonary Arterial Hypertension. Hypertension (Dallas, Tex.: 1979), 79(8), 1580–1590. https://doi.org/10.1161/HYPERTENSIONAHA.122.18261

9. Guignabert, C., & Dorfmuller, P. (2013). Pathology and pathobiology of pulmonary hypertension. Seminars in Respiratory and Critical Care Medicine, 34(5), 551–559. https://doi.org/10.1055/s-0033-1356496

10. Piper, B., Bogamuwa, S., Hossain, T., Farkas, D., Rosas, L., Green, A. C., Newcomb, G., Sun, N., Ovando-Ricardez, J. A., Horowitz, J. C., Bhagwani, A. R., Yang, H., Kudryashova, T. V., Rojas, M., Mora, A. L., Yan, P., Mallampalli, R. K., Goncharova, E. A., Eckmann, D. M., & Farkas, L. (2024). RAB7 deficiency impairs pulmonary artery endothelial function and promotes pulmonary hypertension. The Journal of Clinical Investigation, 134(3), e169441. https://doi.org/10.1172/JCI169441

11. Yu, Q., & Chan, S. Y. (2017). Mitochondrial and Metabolic Drivers of Pulmonary Vascular Endothelial Dysfunction in Pulmonary Hypertension. Advances in Experimental Medicine and Biology, 967, 373–383. https://doi.org/10.1007/978-3-319-63245-2_24

12. Townsley, M. I. (2012). Structure and composition of pulmonary arteries, capillaries, and veins. Comprehensive Physiology, 2(1), 675–709. https://doi.org/10.1002/cphy.c100081

13. Duong, H. T., Comhair, S. A., Aldred, M. A., Mavrakis, L., Savasky, B. M., Erzurum, S. C., & Asosingh, K. (2011). Pulmonary artery endothelium resident endothelial colony-forming cells in pulmonary arterial hypertension. Pulmonary Circulation, 1(4), 475–486. https://doi.org/10.4103/2045-8932.93547

14. King, J., Hamil, T., Creighton, J., Wu, S., Bhat, P., McDonald, F., & Stevens, T. (2004). Structural and functional characteristics of lung macro- and microvascular endothelial cell phenotypes. Microvascular Research, 67(2), 139–151. https://doi.org/10.1016/j.mvr.2003.11.006

15. Passineau, M. J., Gallo, P. H., Williams, G., Perez, R., & Benza, R. L. (2019). Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization. Journal of Visualized Experiments, 143, 58353. https://doi.org/10.3791/58353

16. Ventetuolo, C. E., Aliotta, J. M., Braza, J., Chichger, H., Dooner, M., McGuirl, D., Mullin, C. J., Newton, J., Pereira, M., Princiotto, A., Quesenberry, P. J., Walsh, T., Whittenhall, M., Klinger, J. R., & Harrington, E. O. (2020). Culture of pulmonary artery endothelial cells from pulmonary artery catheter balloon tips: Considerations for use in pulmonary vascular disease. The European Respiratory Journal, 55(3), 1901313. https://doi.org/10.1183/13993003.01313-2019

17. Jaffe, E. A., Nachman, R. L., Becker, C. G., & Minick, C. R. (1973). Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. The Journal of Clinical Investigation, 52(11), 2745–2756. https://doi.org/10.1172/JCI107470

18. Bashore, T. M., Balter, S., Barac, A., Byrne, J. G., Cavendish, J. J., Chambers, C. E., Hermiller, J. B., Kinlay, S., Landzberg, J. S., Laskey, W. K., McKay, C. R., Miller, J. M., Moliterno, D. J., Moore, J. W. M., Oliver-McNeil, S. M., Popma, J. J., & Tommaso, C. L. (2012). 2012 American College of Cardiology Foundation/Society for Cardiovascular Angiography and Interventions Expert Consensus Document on Cardiac Catheterization Laboratory Standards Update. Journal of the American College of Cardiology, 59(24), 2221–2305. https://doi.org/10.1016/j.jacc.2012.02.010

19. Hoeper, M. M., Bogaard, H. J., Condliffe, R., Frantz, R., Khanna, D., Kurzyna, M., Langleben, D., Manes, A., Satoh, T., Torres, F., Wilkins, M. R., & Badesch, D. B. (2013). Definitions and diagnosis of pulmonary hypertension. Journal of the American College of Cardiology, 62(25 Suppl), D42-50. https://doi.org/10.1016/j.jacc.2013.10.032

20. Maron, B. A. (2023). Revised Definition of Pulmonary Hypertension and Approach to Management: A Clinical Primer. Journal of the American Heart Association, 12(8), e029024. https://doi.org/10.1161/JAHA.122.029024

21. Sharma, D., Shah, R. J., Sreenivasan, J., Kafle, P., Gupta, R., Levine, A., Lanier, G. M., & Aronow, W. S. (2022). The role of serial right heart catheterization in risk stratification and management of pulmonary arterial hypertension. Expert Review of Cardiovascular Therapy, 20(7), 543–547. https://doi.org/10.1080/14779072.2022.2092095

22. Vonk Noordegraaf, A., Chin, K. M., Haddad, F., Hassoun, P. M., Hemnes, A. R., Hopkins, S. R., Kawut, S. M., Langleben, D., Lumens, J., & Naeije, R. (2019). Pathophysiology of the right ventricle and of the pulmonary circulation in pulmonary hypertension: An update. The European Respiratory Journal, 53(1), 1801900. https://doi.org/10.1183/13993003.01900-2018

23. Kutikhin, A. G., Tupikin, A. E., Matveeva, V. G., Shishkova, D. K., Antonova, L. V., Kabilov, M. R., & Velikanova, E. A. (2020). Human Peripheral Blood-Derived Endothelial Colony-Forming Cells Are Highly Similar to Mature Vascular Endothelial Cells yet Demonstrate a Transitional Transcriptomic Signature. Cells, 9(4), 876. https://doi.org/10.3390/cells9040876

24. Matveeva, V., Khanova, M., Sardin, E., Antonova, L., & Barbarash, O. (2018). Endovascular Interventions Permit Isolation of Endothelial Colony-Forming Cells from Peripheral Blood. International Journal of Molecular Sciences, 19(11), 3453. https://doi.org/10.3390/ijms19113453

25.