The effect of fresh frozen plasma (FFP) and human platelet lysate (HPL) on the proliferation rate of human mesenchymal stem cells (MSCs) derived from adipose tissue and umbilical cord blood

authors:

avatar Sina Sadrifar , avatar Saiedeh Erfanian ORCID , avatar Rasoul Baharlou , *


how to cite: Sadrifar S, Erfanian S, Baharlou R. The effect of fresh frozen plasma (FFP) and human platelet lysate (HPL) on the proliferation rate of human mesenchymal stem cells (MSCs) derived from adipose tissue and umbilical cord blood. koomesh. 2023;25(3):e152837. 

Abstract

Introduction: Mesenchymal stem cells (MSCs) are cells that can replicate and differentiate into cells of multiple organs. However, there are challenges regarding their cultivation conditions. Fetal bovine serum (FBS) is the most commonly used culture medium additive for in vitro cultures, despite its undefined composition, potential immunogenicity, and possible prion/zoonotic transmission. For these reasons, significant efforts have been targeted at finding substitutes, such as serum-free media or human platelet lysates (HPL) and fresh frozen plasma (FFP). This study aimed to investigate the effects of the simultaneous use of FFP, HPL, and FBS on the proliferation of MSCs derived from umbilical cord blood and adipose tissues. Materials and Methods: In this study, five platelet concentrate bags were obtained from the Iranian Blood Transfusion Organization, the branch of Jahrom. Relatively, the Platelet lysate was prepared by repeated freezing/thawing method, and dead cells were separated from platelet lysate by centrifugation. MSCs were isolated from the adipose tissue of five patients undergoing hernia surgery or cesarean, and from the umbilical cord of 10 newborns. The cells were cultured in control (FBS) and experimental (FBS+FFP, FFP+HPL and FBS+FFP+HPL) groups. Cells of MSC phenotype were confirmed by flow cytometric analysis and viability was examined using the MTT assay. Results: The results showed that the proliferation and viability of mesenchymal stem cells were increased significantly in the FBS+FFP, FBS+HPL, FFP+HPL, and FBS+FFP+HPL groups compared to the FBS group. Conclusion: In this study, it was shown that FBS+FFP, FBS+HPL, FFP+HPL, and FBS+FFP+HPL had a beneficial effect on the growth and proliferation of MSCs. It seems these biological effects are due to the presence of growth factors and cytokines produced by blood cells. This study confirms that FFP+HPL as a supplement in some culture media may be considered a suitable alternative instead of FBS for the MSC culture media.

References

  • 1.

    Friedenstein AJ. Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method. Exp Hematol 1974; 2: 83-92.

  • 2.

    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143-147.

  • 3.

    Pevsner-Fischer M, Levin S, Zipori D. The origins of mesenchymal stromal cell heterogeneity. Stem Cell Rev Rep 2011; 7: 560-568.

  • 4.

    Patel SA, Sherman L, Munoz J, Rameshwar P. Immunological properties of mesenchymal stem cells and clinical implications. Arch Immunol Ther Exp 2008; 56: 1-8.

  • 5.

    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006; 8: 315-317.

  • 6.

    McPherson RA, Pincus MR. Henry's clinical diagnosis and management by laboratory methods E-book: Elsevier Health Sci 2021.

  • 7.

    Elder S, Thomason J. Effect of platelet-rich plasma on chondrogenic differentiation in three-dimensional culture. Open Orthop J 2014; 8: 78.

  • 8.

    Azouna NB, Jenhani F, Regaya Z, Berraeis L, Othman TB, Ducrocq E, Domenech J. 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 Res Ther 2012; 3: 1-14.

  • 9.

    Rauch C, Feifel E, Amann EM, Sptl HP, Schennach H, Pfaller W, Gstraunthaler G. Alternatives to the use of fetal bovine serum: human platelet lysates as a serum substitute in cell culture media. ALTEX 2011; 28: 305-316.

  • 10.

    Hartwig D, Herminghaus P, Wedel T, Liu L, Schlenke P, Dibbelt L, Geerling G. Topical treatment of ocular surface defects: comparison of the epitheliotrophic capacity of fresh frozen plasma and serum on corneal epithelial cells in an in vitro cell culture model. Transfus Med 2005; 15: 107-113.

  • 11.

    Arpornmaeklong P, Kochel M, Depprich R, Kbler N, Wrzler K. Influence of platelet-rich plasma (PRP) on osteogenic differentiation of rat bone marrow stromal cells. An in vitro study. Int J Oral Maxillofac Surg 2004; 33: 60-70.

  • 12.

    Schallmoser K, Strunk D. Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures. J Vis Exp 200; 1e1523.

  • 13.

    Xia W, Li H, Wang Z, Xu R, Fu Y, Zhang X, et al. Human platelet lysate supports ex vivo expansion and enhances osteogenic differentiation of human bone marrowderived mesenchymal stem cells. Cell Biol Int 2011; 35: 639-643.

  • 14.

    Fazzina R, Iudicone P, Mariotti A, Fioravanti D, Procoli A, Cicchetti E, et al. Culture of human cell lines by a pathogen-inactivated human platelet lysate. Cytotechnology 2016; 68: 1185-1195.

  • 15.

    Schallmoser K, Bartmann C, Rohde E, Reinisch A, Kashofer K, Stadelmeyer E, et al. Human platelet lysate can replace fetal bovine serum for clinicalscale expansion of functional mesenchymal stromal cells. Transfusion 2007; 47: 1436-146.

  • 16.

    Dessels C, Potgieter M, Pepper MS. Making the switch: alternatives to fetal bovine serum for adipose-derived stromal cell expansion. Front Cell Dev Biol 2016; 4: 115.

  • 17.

    Baharlou R, Tajik N, Behdani M, Shokrgozar MA, Tavana V, et al. An antibody fragment against human delta-like ligand-4 for inhibition of cell proliferation and neovascularization. Immunopharmacol Immunotoxicol 2018; 40: 368-374.

  • 18.

    Mohammadlou M, Salehi S, Baharlou R. Development of anti DLL4 Nanobody fused to truncated form of Pseudomonas exotoxin: As a novel immunotoxin to inhibit of cell proliferation and neovascularization. Anal Biochem 2022; 653: 114776.

  • 19.

    Chevallier N, Anagnostou F, Zilber S, Bodivit G, Maurin S, Barrault A, et al. Osteoblastic differentiation of human mesenchymal stem cells with platelet lysate. Biomaterials 2010; 31: 270-278.

  • 20.

    Amir Allahverdi, Arefeh Jafarian, Saeed Abroun, Masoud Soleimani, Mohammad Taghikhani, Fatemeh Eskandari. The effect of platelet lysate on proliferation and differentiation of mesenchymal stem cells. Pajoohande 2014; 19: 125-30.

  • 21.

    Rahimi Mofrad M, Yari F, Nikougoftar Zarif M, Dadashi M, Aghaie A. Comparison of the effect of platelet lysate derived from umbilical cord blood and peripheral blood on the expansion and differentiation of MSCs. Scientific Journal of Iran Blood Transfus Organ 2021; 18: 255-68.

  • 22.

    Avanzini MA, Bernardo ME, Cometa AM, Perotti C, Zaffaroni N, Novara F, et al. Generation of mesenchymal stromal cells in the presence of platelet lysate: a phenotypic and functional comparison of umbilical cord blood-and bone marrow-derived progenitors. Haematologica 2009; 94: 1649.

  • 23.

    Lv FJ, Tuan RS, Cheung KM, Leung VY. Concise review: the surface markers and identity of human mesenchymal stem cells. Stem Cells 2014; 32: 1408-1419.

  • 24.

    Zhang ZY, Teoh SH, Hui JH, Fisk NM, Choolani M, Chan JK. The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application. Biomaterials 2012; 33: 2656-2672.

  • 25.

    Jonsdottir-Buch SM, Lieder R, Sigurjonsson OE. Platelet lysates produced from expired platelet concentrates support growth and osteogenic differentiation of mesenchymal stem cells. PloS One 2013; 8: e68984.

  • 26.

    Crespo-Diaz R, Behfar A, Butler GW, Padley DJ, Sarr MG, Bartunek J, et al. Platelet lysate consisting of a natural repair proteome supports human mesenchymal stem cell proliferation and chromosomal stability. Cell Transplant 2011; 20: 797-812.

  • 27.

    Astori G, Amati E, Bambi F, Bernardi M, Chieregato K, Schfer R, et al. Platelet lysate as a substitute for animal serum for the ex-vivo expansion of mesenchymal stem/stromal cells: present and future. Stem Cell Res Ther 2016; 7: 1-8.

  • 28.

    Viau S, Lagrange A, Chabrand L, Lorant J, Charrier M, Rouger K, et al. A highly standardized and characterized human platelet lysate for efficient and reproducible expansion of human bone marrow mesenchymal stromal cells. Cytotherapy 2019; 21: 738-754.

  • 29.

    Samuel S, Ahmad RE, Ramasamy TS, Karunanithi P, Naveen SV, Murali MR, et al. Platelet-rich concentrate in serum free medium enhances osteogenic differentiation of bone marrow-derived human mesenchymal stromal cells. PeerJ 2016; 4: e2347.

  • 30.

    Stessuk T, Puzzi MB, Chaim EA, Alves PCM, de Paula EV, Forte A, Izumizawa JM, et al. Platelet-rich plasma (PRP) and adipose-derived mesenchymal stem cells: stimulatory effects on proliferation and migration of fibroblasts and keratinocytes in vitro. Arch Dermatol Res 2016; 308: 511-520.

  • 31.

    Shanbhag S, Mohamed-Ahmed S, Lunde THF, Suliman S, Bolstad AI, Hervig T, Mustafa K. Influence of platelet storage time on human platelet lysates and platelet lysate-expanded mesenchymal stromal cells for bone tissue engineering. Stem Cell Res Ther 2020; 11: 1-18.

  • 32.

    Mareschi K, Castiglia S, Adamini A, Rustichelli D, Marini E, Banche Niclot AG, et al. Inactivated platelet lysate supports the proliferation and immunomodulant characteristics of mesenchymal stromal cells in GMP culture conditions. Biomedicines 2020; 8: 220.

  • 33.

    Kinzebach S, Dietz L, Klter H, Thierse HJ, Bieback K. Functional and differential proteomic analyses to identify platelet derived factors affecting ex vivo expansion of mesenchymal stromal cells. BMC Cell Biol 2013; 14: 1-13.

  • 34.

    Hemeda H, Giebel B, Wagner W. Evaluation of human platelet lysate versus fetal bovine serum for culture of mesenchymal stromal cells. Cytotherapy 2014; 16: 170-180.