Optimization of Animal Sera-Free Culture Condition for Generation and Expansion of Human Cardiosphere-Derived Cells
Background: Preclinical studies have introduced cardiac stem/progenitor cells (CSCs) as a promising cell candidate for cell therapy of heart diseases. CSCs can be isolated from myocardial biopsies using various protocols, expanded in vitro and transplanted back to the patients. One of the most important issues regarding the clinical usage of cells is the choice of suitable humanized culture supplements to replace commonly used animal-derived products including fetal bovine serum (FBS) or fetal calf serum (FCS).
Methods and Materials: In order to find the optimal FBS substitute, human myocardial samples were cultured as explants in media supplemented with one of these different blood products: FBS, human serum (HS), human plasma (HP) or platelet lysate (PL). The out-grown cells were cultured in suspension to generate cardiospheres and then plated to expand as cardiosphere-derived cells (CDCs). The effect of culture media on the process of CDC generation and culture was evaluated in terms of morphology and cell growth.
Results: Among the examined humanized agents, CDCs were only generated and expanded in medium supplemented with HS. Furthermore, they had normal karyotype and expressed CSC associated surface markers but not endothelial and hematopoietic markers. Moreover, cultured CDCs in HS inhibited the proliferation of induced lymphocytes in vitro which might highlight the immuno-modulatory feature of these cells.
Conclusion: Taken together, our data exhibited the superiority of HS compared to other tested human blood products for CDC culture which can be suggested for cell culture set up of cardiac clinical studies.
2. Gersh BJ, Sliwa K, Mayosi BM, Yusuf S. Novel therapeutic concepts: the epidemic of cardiovascular disease in the developing world: global implications. European heart journal. 2010;31(6):642-8.
3. Roth GA, Johnson C, Abajobir A, Abd-Allah F, Abera SF, Abyu G, et al. Global, Regional, and National Burden of Cardiovascular Diseases for 10 Causes, 1990 to 2015. Journal of the American College of Cardiology. 2017;70(1):1-25.
4. Oh H, Ito H, Sano S. Challenges to success in heart failure: Cardiac cell therapies in patients with heart diseases. Journal of cardiology. 2016;68(5):361-7.
5. Chen C, Termglinchan V, Karakikes I. Concise Review: Mending a Broken Heart: The Evolution of Biological Therapeutics. Stem cells. 2017;35(5):1131-40.
6. Cho GS, Fernandez L, Kwon C. Regenerative medicine for the heart: perspectives on stem-cell therapy. Antioxidants & redox signaling. 2014;21(14):2018-31.
7. Sanz-Ruiz R, Fernandez-Aviles F. Autologous and allogeneic cardiac stem cell therapy for cardiovascular diseases. Pharmacological research. 2018;127:92-100.
8. Bianconi V, Sahebkar A, Kovanen P, Bagaglia F, Ricciuti B, Calabro P, et al. Endothelial and cardiac progenitor cells for cardiovascular repair: A controversial paradigm in cell therapy. Pharmacology & therapeutics. 2018;181:156-68.
9. Vahdat S, Mousavi SA, Omrani G, Gholampour M, Sotoodehnejadnematalahi F, Ghazizadeh Z, et al. Cellular and molecular characterization of human cardiac stem cells reveals key features essential for their function and safety. Stem cells and development. 2015;24(12):1390-404.
10. Li TS, Cheng K, Lee ST, Matsushita S, Davis D, Malliaras K, 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.
11. Davis DR, Kizana E, Terrovitis J, Barth AS, Zhang Y, Smith RR, et al. Isolation and expansion of functionally-competent cardiac progenitor cells directly from heart biopsies. Journal of molecular and cellular cardiology. 2010;49(2):312-21.
12. Messina E, De Angelis L, Frati G, Morrone S, Chimenti S, Fiordaliso F, et al. Isolation and expansion of adult cardiac stem cells from human and murine heart. Circulation research. 2004;95(9):911-21.
13. Usta SN, Scharer CD, Xu J, Frey TK, Nash RJ. Chemically defined serum-free and xeno-free media for multiple cell lineages. Annals of translational medicine. 2014;2(10):97.
14. Panchalingam KM, Jung S, Rosenberg L, Behie LA. Bioprocessing strategies for the large-scale production of human mesenchymal stem cells: a review. Stem cell research & therapy. 2015;6:225.
15. Tekkatte C, Gunasingh GP, Cherian KM, Sankaranarayanan K. "Humanized" stem cell culture techniques: the animal serum controversy. Stem cells international. 2011;2011:504723.
16. Karnieli O, Friedner OM, Allickson JG, Zhang N, Jung S, Fiorentini D, et al. A consensus introduction to serum replacements and serum-free media for cellular therapies. Cytotherapy. 2017;19(2):155-69.
17. Angelini F, Ionta V, Rossi F, Miraldi F, Messina E, Giacomello A. Foetal bovine serum-derived exosomes affect yield and phenotype of human cardiac progenitor cell culture. BioImpacts : BI. 2016;6(1):15-24.
18. Monsanto MM, White KS, Kim T, Wang BJ, Fisher K, Ilves K, et al. Concurrent Isolation of 3 Distinct Cardiac Stem Cell Populations From a Single Human Heart Biopsy. Circulation research. 2017;121(2):113-24.
19. Gaetani R, Feyen DA, Doevendans PA, Gremmels H, Forte E, Fledderus JO, et al. Different types of cultured human adult cardiac progenitor cells have a high degree of transcriptome similarity. Journal of cellular and molecular medicine. 2014;18(11):2147-51.
20. Poloni A, Maurizi G, Serrani F, Mancini S, Discepoli G, Tranquilli AL, et al. Human AB serum for generation of mesenchymal stem cells from human chorionic villi: comparison with other source and other media including platelet lysate. Cell proliferation. 2012;45(1):66-75.
21. Tunaitis V, Borutinskaite V, Navakauskiene R, Treigyte G, Unguryte A, Aldonyte R, et al. Effects of different sera on adipose tissue-derived mesenchymal stromal cells. Journal of tissue engineering and regenerative medicine. 2011;5(9):733-46.
22. Kocaoemer A, Kern S, Kluter H, Bieback K. Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue. Stem cells. 2007;25(5):1270-8.
23. Ben Azouna N, Jenhani F, Regaya Z, Berraeis L, Ben Othman T, Ducrocq E, et al. Phenotypical and functional characteristics of mesenchymal stem cells from bone marrow: comparison of culture using different media supplemented with human platelet lysate or fetal bovine serum. Stem cell research & therapy. 2012;3(1):6.
24. Perez-Ilzarbe M, Diez-Campelo M, Aranda P, Tabera S, Lopez T, del Canizo C, et al. Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy. Transfusion. 2009;49(9):1901-10.
25. Juhl M, Tratwal J, Follin B, Sondergaard RH, Kirchhoff M, Ekblond A, et al. Comparison of clinical grade human platelet lysates for cultivation of mesenchymal stromal cells from bone marrow and adipose tissue. Scandinavian journal of clinical and laboratory investigation. 2016;76(2):93-104.
26. Siciliano C, Chimenti I, Bordin A, Ponti D, Iudicone P, Peruzzi M, et al. The potential of GMP-compliant platelet lysate to induce a permissive state for cardiovascular transdifferentiation in human mediastinal adipose tissue-derived mesenchymal stem cells. BioMed research international. 2015;2015:162439.
27. Yoon AY, Yun S, Yang H, Lim YH, Kim H. Expression of tight junction molecule in the human serum-induced aggregation of human abdominal adipose-derived stem cells in vitro. Development & reproduction. 2014;18(4):213-24.
28. Tateishi K, Ando W, Higuchi C, Hart DA, Hashimoto J, Nakata K, et al. Comparison of human serum with fetal bovine serum for expansion and differentiation of human synovial MSC: potential feasibility for clinical applications. Cell transplantation. 2008;17(5):549-57.
29. Lindroos B, Aho KL, Kuokkanen H, Raty S, Huhtala H, Lemponen R, et al. Differential gene expression in adipose stem cells cultured in allogeneic human serum versus fetal bovine serum. Tissue engineering Part A. 2010;16(7):2281-94.
30. Chimenti I, Gaetani R, Forte E, Angelini F, De Falco E, Zoccai GB, et al. Serum and supplement optimization for EU GMP-compliance in cardiospheres cell culture. Journal of cellular and molecular medicine. 2014;18(4):624-34.
31. Nazari-Shafti TZ, Xu Z, Bader AM, Henke G, Klose K, Falk V, et al. Mesenchymal Stromal Cells Cultured in Serum from Heart Failure Patients Are More Resistant to Simulated Chronic and Acute Stress. Stem cells international. 2018;2018:5832460.
32. Mizuno M, Katano H, Otabe K, Komori K, Kohno Y, Fujii S, et al. Complete human serum maintains viability and chondrogenic potential of human synovial stem cells: suitable conditions for transplantation. Stem cell research & therapy. 2017;8(1):144.
33. Dos Santos VT, Mizukami A, Orellana MD, Caruso SR, da Silva FB, Traina F, et al. Characterization of Human AB Serum for Mesenchymal Stromal Cell Expansion. Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie. 2017;44(1):11-21.
34. Chachques JC, Herreros J, Trainini J, Juffe A, Rendal E, Prosper F, et al. Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular cardiomyoplasty. International journal of cardiology. 2004;95 Suppl 1:S29-33.
35. Ferro F, Spelat R, Beltrami AP, Cesselli D, Curcio F. Isolation and characterization of human dental pulp derived stem cells by using media containing low human serum percentage as clinical grade substitutes for bovine serum. PloS one. 2012;7(11):e48945.