Comparative study of the effect of bFGF and plasma rich in growth factors on cryopreserved multipotent mesenchymal stromal cells from bone marrow and tendon of rats
Volkovа N. A., Yukhta M. S., Goltsev A. N.
Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine
The purpose of study was to investigate in vitro effects of growth factors, known as cell proliferation stimulants, to determine the most suitable agent for enhancing the proliferation and migration activity of cryopreserved multipotent mesenchymal stromal cells (MMSCs) derived from bone marrow and tendon tissue.
Materials and methods. MMSCs were obtained from bone marrow and tendon tissues of rats. Cryopreservation was carried out under the protection of 10 % DMSO with the addition of 20 % fetal bovine serum at a cooling rate of 1°C/min to -80°C and subsequent freeze in liquid nitrogen. During the cultivation of the cryopreserved MMSCs, basis fibroblast growth factor (bFGF) and plasma rich in growth factors were used. The ability to proliferation (MTT assay), migration (in vitro scratch assay), and the synthesis of collagen type I (immunocytochemical study of collagen type I expression) were evaluated.
Results. The use of plasma rich in growth factors contributes to increasing the ability of cryopreserved MMSCs from bone marrow to proliferate and migrate, associated with decreasing in the relative number of cells that express collagen type I. Cultures of cryopreserved MMSCs from the tendon tissue exhibit greater sensitivity to the bFGF compared to the plasma rich in growth factors that have a manifestation in the increasing of cell proliferation and migration ability.
Conclusions. bFGF and plasma rich in growth factors can be used as stimulants for stromal cell cultures.
Keywords: multipotent mesenchymal stromal cells; bone marrow; tendon tissue; basic fibroblast growth factor; plasma rich in growth factorsFull Text PDF
|1. Tozer S, Duprez D. Tendon and ligament: development, repair and disease. Birth Defects Res C Embryo Today. 2005; 75(3): 226-236.
|2. Kyryk VM, Butenko GM. Stvolovye kletki iz zhirovoy tkani: osnovnye kharakteristiki i perspektivy klinicheskogo primeneniya v regenerativnoy meditsine (obzor literatury) [Stem cells from adipose tissue: the main characteristics and perspectives of clinical use in regenerative medicine (review)]. Zhurnal Akademії medichnikh nauk Ukraїni – Journal of the Academy of Medical Sciences of Ukraine. 2010; 16(4): 576-604. [In Russian]|
|3. Volkova N, Yukhta M, Goltsev A. Cryopreserved Mesenchymal Stem Cells Stimulate Regeneration in an Intervertebral Disc. Biomedicines. 2015; 3(3): 237-247.
|4. Rybachuk ОА, Кyryk VМ, Poberezhny PA, et al. Еffect of the bone marrow multipotent mesenchimal stromal cells to the neural tissue after ischemic injury in vitro. Cell and Organ Transplantology. 2014; 2(1): 74-78.
|5. Volkova NА, Yukhta MS, Yurchuk ТА, et al. Multipotent mesenchymal stromal cells of bone marrow in therapy of chronic inflammation of murine ovaries. Biotechnologia Acta. 2014; 7(5): 35-42.
|6. Ra J, Kang S, Shin S. Stem cell treatment for patients with autoimmune disease by systemic infusion of culture expanded autologous adipose tissue derived mesenchymal stem cells. J Transl Med. 2011; 9(1): 181-198.|
|7. Hegyi B, Sagi1 B, Kovacs J. Identical, similar or different? Learning about immunomodulatory function of mesenchymal stem cells isolated from various mouse tissues: bone marrow, spleen, thymus and aorta wall. Int Immunology. 2010; 22(7): 551-559.
|8. Graham R. Tendinopathy – from basic science to treatment. Nature Clinical Practice Rheumatology. 2008; 4(2): 82-89.
|9. Du M, Zhu T, Duan X, et al. Acellular dermal matrix loading with bFGF achieves similar acceleration of bone regeneration to BMP-2 via differential effects on recruitment, proliferation and sustained osteodifferentiation of mesenchymal stem cells. Mater Sci Eng C Mater Biol Appl. 2016; 70(1): 62-70.|
|10. Bocelli-Tyndall C, Zajac P, Maggio N, et al. Fibroblast growth factor 2 and platelet-derived growth factor, but not platelet lysate, induce proliferation-dependent, functional class II major histocompatibility complex antigen in human mesenchymal stem cells. Arthritis Rheum. 2010; 62(12): 3815-3825.
|11. Chan BP, Fu SC, Qin L, et al. Effects of basic fibroblast growth factor (bFGF) on early stages of tendon healing: a rat patellar tendon model. Acta Orthopaedica Scandinavica. 2000; 71(5): 513-518.
|12. Molloy T, Wang Y, Murrell GAC, Molloy T. The roles of growth factors in tendon and ligament healing. Sports Medicine. 2003; 33(5): 381-394.
|13. Ehrenfest D, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009; 27(3): 158-167.
|14. Ehrenfest D, Bielecki T, Mishra A, et al. In search of a consensus terminology in the field of platelet concentrates for surgical use: platelet-rich plasma (PRP), platelet-rich fibrin (PRF), fibrin gel polymerization and leukocytes). Curr Pharm Biotechnol. 2012; 13(7): 1131-1137.
|15. Anitua E, Sanchez M, Merayo-Lloves J, et al. Plasma rich in growth factors (PRGF-Endoret) stimulates proliferation and migration of primary keratocytes and conjunctival fibroblasts and inhibits and reverts TGF-β1–induced myodifferentiation). Investigative ophthalmology & visual science. 2011; 52(9): 6066-6073.
|16. Nishiyama K, Okudera T, Watanabe T, et al. Basic characteristics of plasma rich in growth factors (PRGF): blood cell components and biological effects). Clin Exp Dent Res. 2016; 2(2): 96-103.
|17. Anitua E. Plasma rich in growth factors: preliminary results of use in the preparation of future sites for implants. International journal of Oral and maxillofacial Implants. 1999; 14(4): 529-535.|
|18. Sanchez M, Anitua E, Azofra J, et al. Comparison of surgically repaired Achilles tendon tears using platelet-rich fibrin matrices. Am J Sports Med. 2007; 35(2): 245–251.
|19. Sаnchez M, Anitua E, Azofra J, et al. Intra-articular injection of an autologous preparation rich in growth factors for the treatment of knee OA: a retrospective cohort study. Clin Exp Rheumatol. 2008; 269(5): 910-913|
|20. Costa MA, Wu C, Pham BV, et al. Tissue engineering of flexor tendons: optimization of tenocyte proliferation using growth factor supplementation. Tissue engineering. 2006; 12(7): 1937-1943.
|21. Anitua E, Sаnchez M, Orive G, et al. The potential impact of the preparation rich in growth factors (PRGF) in different medical fields. Biomaterials. 2007; 28(31): 4551-4560.
|22. Anitua E, Pino A, Orive G. Plasma rich in growth factors promotes dermal fibroblast proliferation, migration and biosynthetic activity. J Wound Care. 2016; 25(11): 680-687.
|23. Volkovа NA, Yukhta MS, Goltsev AN. Morphological and functional characteristics of cryopreserved multipotent mesenchymal stromal cells from bone marrow, adipose tissue and tendons. Cell and Organ Transplantology. 2016; 4(2): 200-205.
|24. Council of Europe [France]. European convention for the protection of vertebrate animals used for experimental and other scientific purposes. Strasbourg, 18.III.1986, http://conventions.coe.int/treaty/en/Treaties/Word/123.doc|
|25. Volkova NA, Goltsev AN. Сryopreservation effect on proliferation and differentiation potential of cultured chorion cells. CryoLetters. 2015; 36(1): 25-29.|
|26. Song G, Ju Y, Soyama H. Growth and proliferation of bone marrow mesenchymal stem cells affected by type I collagen, fibronectin and bFGF. Materials Science and Engineering: 2008; 28(8): 1467-1471.
|27. Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2): 55-63.|
|28. Vigano M, Orfei CP, Colombini A, et al. Different culture conditions affect the growth of human tendon stem/progenitor cells (TSPCs) within a mixed tendon cells (TCs) population. Journal of experimental orthopedics. 2017; 4(1): 8.
|29. Maffulli N, Ewen SW, Waterston SW, et al. Tenocytes from ruptured and tendinopathic achilles tendons produce greater quantities of type III collagen than tenocytes from normal achilles tendons: an in vitro model of human tendon healing. Am J Sports Med. 2000; 28(4): 499-505.
|30. Chan BP, Fu S, Qin L, et al. Effect of basic fibroblast growth factor (bFGF) on early stages of tendon healing: a rat patellar tendon model. Acta Orthop Scand. 2000; 71: 513.
|31. Folkman J, Klagsbrun M. Angiogenic factors. Science. 1987; 235(4787): 442-447.
|32. Anitua E, Sanchez M, de la Fuente M, et al. Plasma rich in growth factors (PRGF-Endoret) stimulates tendon and synovial fibroblasts migration and improves the biological properties of hyaluronic acid. Knee Surg Sports Traumatol Arthrosc. 2012; 20(9): 1657-1665.
|33. Anitua E, Troya M, Orive G. Plasma rich in growth factors promote gingival tissue regeneration by stimulating fibroblast proliferation and migration and by blocking transforming growth factor-β1-induced myodifferentiation. Orive Periodontol. 2012; 83(8): 1028-1037.
|34. Anitua E, Tejero R, Zalduendo MM, et al. Plasma rich in growth factors promotes bone tissue regeneration by stimulating proliferation, migration, and autocrine secretion in primary human osteoblasts. J Periodontol. 2013; 84(8): 1180-1190.
Volkovа NA, Yukhta MS, Goltsev AN. Comparative study of the effect of bFGF and plasma rich in growth factors on cryopreserved multipotent mesenchymal stromal cells from bone marrow and tendon of rats. Cell and Organ Transplantology. 2017; 5(2):170-175. doi:10.22494/cot.v5i2.75