Bioluminescent monitoring of rat cardiosphere-derived cells in platelet gel engraftment in ischemic heart

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access


The success of cell therapy depends on an effective method of cell delivery and engraftment. The use of transgenic cells carrying a reporter system based on the luciferase gene allows to perform the quantitative evaluation of the transplantation efficiency in dynamics using biochemical methods. The purpose of this work was to monitor the persistence of rat cardiosphere-derived cells (CDC) after allogeneic transplantation into the periinfarction zone. Transplantation was performed by intramyocardial injection of a cell suspension in a culture medium or in platelet rich plasma (PRP). When injected into the myocardium PRP forms fibrin clots which serves as a matrix for the transplanted cells. The cells were modified by the luciferase enzyme gene by transduction with lentiviruses (CDC-Luc). The activity of luciferase was determined in protein extracts of the myocardium at different time points after the transplantation. It was shown that in the first hour after injections, CDC-Luc is quantitatively detected in the peri-infarction zone irrespective of the use of platelet gel or medium, and their amount does not decrease within 48 hours. During this period, we found a positive effect of the fibrin matrix on the cells - the luminescence of CDC-Luc protein extracts in the platelet gel composition was significantly higher. We suggested that the platelet gel promotes a more favorable microenvironment for CDC-Luc and facilitates the adaptation of cells after transplantation, what reflected in the recovery of the level of luciferase production in cells. Further, we found negative dynamics: CDC-Luc injected in the culture medium is retained in the myocardium for 5 days and on the seventh day their presence is not determined, CDC-Luc in the fibrin matrix is retained in the myocardium for 10 days after transplantation. Thus, despite the successful transplantation of CDC, the integration of cells into the myocardium does not occur. Nevertheless, the use of platelet gel prolongates the time of CDC persistence in the tissue and enhances of their paracrine effect. The use of fibrin matrix can be useful for long-lived cells, such as cardiomyocytes, in particular, to improve the efficiency of transplantation of the tissue engineering biological pacemaker. A luciferase reportering system can be effective for in vitro and in vivo monitoring of cell fate, both in biotechnological stages of cultivation and assembly of the tissue engineering biopee maker, and after myocardial transplantation. In the future, the developed methodological approach will be used to study of tissue-engineering biopacemakers in experimental animals.

Full Text

Restricted Access

About the authors

S. V Pavlova

Institute of Cytology and Genetics of Siberian Branch of the RAS; E.N. Meshalkin National Medical Research Center; Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the RAS


E. A Leonova

E.N. Meshalkin National Medical Research Center

E. V Chepeleva

E.N. Meshalkin National Medical Research Center

A. A Dokuchaeva

E.N. Meshalkin National Medical Research Center

D. S Sergeevichev

E.N. Meshalkin National Medical Research Center

E. A Pokushalov

E.N. Meshalkin National Medical Research Center


  1. Gyongyosi M., Wojakowski W., Lemarchand P. et al. Meta-Analysis of Cell-based CaRdiac stUdiEs [ACCRUE) in Patients With Acute Myocardial Infarction Based on Individual Patient Data. Circ. Res. 2015; 116(8): 1346-60.
  2. Rossini A., Frati C., Lagrasta C. et al. Human cardiac and bone marrow stromal cells exhibit distinctive properties related to their origin. Cardiovasc. Res. 2011; 89(3): 650-60.
  3. Bolli R., Chugh A.R., D'Amario D. et al. Cardiac stem cells in patients with ischaemic cardiomyopathy (SCIPIO): initial results of a randomised phase 1 trial. Lancet 2011; 378(9806): 1847-57.
  4. Makkar R.R., Smith R.R., Cheng K. et al. Intracoronary cardiosphere-derived cells for heart regeneration after myocardial infarction (CADUCEUS): a prospective, randomised phase 1 trial. Lancet 2012; 379(9819): 895-904.
  5. Malliaras K., Makkar R.R., Smith R.R. et al. Intracoronary Cardiosphere-Derived Cells After Myocardial Infarction. J. Am. Coll. Cardiol. 2014; 63(2): 110-22.
  6. Birket M.J., Ribeiro M.C., Verkerk A.O. et al. Expansion and patterning of cardiovascular progenitors derived from human pluripotent stem cells. Nat. Biotechnol. 2015; 33(9): 970-9.
  7. Chong J.H., Yang X., Don C.W. et al. Human embryonic-stemcell-derived cardiomyocytes regenerate non-human primate hearts. Nature 2014; 510(7504): 273-7.
  8. Husse B., Franz W.M. Generation of cardiac pacemaker cells by programming and differentiation. Biochim. Biophys. Acta - Mol. Cell Res. 2016; 1863(7): 1948-52.
  9. Chepeleva E.V., Pavlova S.V., Malakhova A.A. et al. Therapy of Chronic Cardiosclerosis in WAG Rats Using Cultures of Cardiovascular Cells Enriched with Cardiac Stem Cells. Bull. Exp. Biol. Med. 2015; 160(1): 165-73.
  10. Terrovitis J.V., Smith R.R., Marban E. Assessment and Optimization of Cell Engraftment After Transplantation Into the Heart. Circ. Res. 2010; 106(3): 479-94.
  11. Cheng K., Malliaras K., Shen D. et al. Intramyocardial injection of platelet gel promotes endogenous repair and augments cardiac function in rats with myocardial infarction. J. Am. Coll. Cardiol. 2012; 59(3): 256-64.
  12. Smith A.J., Lewis F.C., Aquila I. et al. Isolation and characterization of resident endogenous c-Kit+ cardiac stem cells from the adult mouse and rat heart. Nat. Protoc. 2014; 9(7): 1662-81.
  13. Messina E., De Angelis L., Frati G. et al. Isolation and Expansion of Adult Cardiac Stem Cells From Human and Murine Heart. Circ. Res. 2004; 95(9): 911-21.
  14. Pavlova S.V., Perovskii P.P., Chepeleva E.V. et al. Characteristics of cardiac cell cultures derived from human myocardial explants. Bull. Exp. Biol. Med. 2013; 156(1): 127-35.
  15. Kingston R.E., Chen C.A., Rose J.K. Calcium Phosphate Transfection. In: Hoboken N.J., editor. Current Protocols in Molecular Biology. USA: John Wiley & Sons Inc.; 2003. р. 9.1.1-11.
  16. Григорьева Е.В., Шевченко А.И., Медведев С.П. и соавт. Индуцированные плюрипотентные стволовые клетки гибридов полёвок Microtus levis * Microtus arvalis : условия, необходимые для получения и поддержания. Acta Naturae 2015; 4(27): 64-78.
  17. Милевская Е.А., Немудрый А.А., Чепелева Е.В. и соавт. Оптимизация протокола интрамиокардиальной трансплантации с использованием люминесценции кардиальных мезенхимальных клеток, маркированных экспрессией люциферазы. Патология кровообращения и кардиохирургия 2015; 19(4-2): 69-76.
  18. Cheng K., Shen D., Smith J. et al. Transplantation of platelet gel spiked with cardiosphere-derived cells boosts structural and functional benefits relative to gel transplantation alone in rats with myocardial infarction. Biomaterials 2012; 33(10): 2872-9.
  19. Chang C., Chan A., Lin X. et al. Cellular bioenergetics is an important determinant of the molecular imaging signal derived from luciferase and the sodium-iodide symporter. Circ. Res. 2013; 112(3): 441-50.
  20. Manthorpe M., Cornefert-Jensen F., Hartikka J. et al. Gene Therapy by Intramuscular Injection of Plasmid DNA: Studies on Firefly Luciferase Gene Expression in Mice. Hum. Gene Ther. 1993; 4(4): 419-31.
  21. Koninckx R., Daniäls A., Windmolders S. et al. Mesenchymal stem cells or cardiac progenitors for cardiac repair? A comparative study. Cell. Mol. Life Sci. 2011; 68(12): 2141-56.
  22. Kasai-Brunswick T.H., Costa A.R., Barbosa R.A. et al. Cardiosphere-derived cells do not improve cardiac function in rats with cardiac failure. Stem Cell Res. Ther. 2017; 8(1): 36.
  23. Павлова С.В., Сергеевичев Д.С., Чепелева Е.В. и др. Сравнение мезенхимальных стромальных клеток костного мозга и региональных стволовых клеток сердца и фибробластов кожи человека. Патология кровообращения и кардиохирургия 2015; 19(4-2): 12-9.
  24. Захарова И.С., Живень М.К., Саая Ш.Б. и соавт. Разработка клеточных технологий для создания клеточно-наполненных сосудистых трансплантатов. Патология кровообращения и кардиохирургия 2015; 19(4-2): 43-54.
  25. Zakharova I.S., Zhiven' M.K., Saaya S.B. et al. Endothelial and smooth muscle cells derived from human cardiac explants demonstrate angiogenic potential and suitable for design of cell-containing vascular grafts. J. Transl. Med. 2017; 15(1): 54.
  26. Andersen D.C., Andersen P., Schneider M. et al. Murine “Cardiospheres” Are Not a Source of Stem Cells with Cardiomyogenic Potential. Stem Cells 2009; 27(7): 1571-81.
  27. Павлова С.В., Розанова И.А., Чепелева Е.В. и др. Ангиогенный потенциал кардиальных стволовых и мезенхимальных стромальных клеток костного мозга крысы. Патология кровообращения и кардиохирургия 2015; 19(4-2): 77-84.
  28. Li T.S., Cheng K., Lee S.T. et al. Cardiospheres Recapitulate a Niche-Like Microenvironment Rich in Stemness and Cell-Matrix Interactions, Rationalizing Their Enhanced Functional Potency for Myocardial Repair. Stem Cells 2010; 28(11): 2088-98.
  29. Keith M.C., Bolli R. "String Theory" of c-kit pos Cardiac Cells. Circ. Res. 2015; 116(7): 1216-30.
  30. van Berlo J.H., Kanisicak O., Maillet M. et al. C-Kit+ Cells Minimally Contribute Cardiomyocytes To the Heart. Nature 2014; 509(7500): 337-41.
  31. Sultana N., Zhang L., Yan J. et al. Resident c-kit(+) cells in the heart are not cardiac stem cells. Nat. Commun. 2015; 6: 8701.
  32. Laflamme M.A., Murry C.E. Regenerating the heart. Nat. Biotechnol. 2005; 23(7): 845-56.
  33. Hong K.U., Guo Y., Li Q.H. et al. c-kit+ Cardiac Stem Cells Alleviate Post-Myocardial Infarction Left Ventricular Dysfunction Despite Poor Engraftment and Negligible Retention in the Recipient Heart. PLoS One 2014; 9(5): e96725.
  34. Johnston P.V., Sasano T., Mills K. et al. Engraftment, differentiation, and functional benefits of autologous cardiospherederived cells in porcine ischemic cardiomyopathy. Circulation 2009; 120(12): 1075-83.

Copyright (c) 2017 Eco-Vector

СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: ПИ № ФС 77 - 85657 от 21.07.2023 от 11.03.2014.

This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies