IN VITRO PROPAGATION OF PLATELETS FROM EMBRYONIC STEM CELLS WITH C. PAPAYA LEAF EXTRACT

Authors

  • ADARSH D. B. Genelon Institute of Life Sciences, Bangalore, India
  • ELANGO E. MURUGAIAN Genelon Institute of Life Sciences, Bangalore, India. Evergreen Biosolutions, HIG 162, Avalapalli Hudco, Hosur-635109 Tamilnadu, India

DOI:

https://doi.org/10.22159/ijcpr.2023v15i4.3024

Keywords:

Platelet, Embryonic stem cells, Megakaryocytes, Thrombopoietin, Papaya leaf extract

Abstract

Objective: The depletion of platelets in patients under different clinical conditions like thrombocytopenia is often associated with a significant mortality factor. Alternative modes of platelet propagation from other resources, including stem cells, are evaluated under in vitro conditions. Embryonic stem (ES) cells can proliferate and differentiate into various cell types. This study evaluated the generation of functional platelets from embryonic stem cells with C. papaya leaf extract through a controlled sequential pathway to meet the clinical demand.

Methods: In vitro culture of embryonic stem cells was carried out with the help of growth factors and a suitable medium. The differentiated megakaryocytes and platelets were analyzed by staining, flow cytometer and functional studies. The normal megakaryocytes were compared with the megakaryocytes differentiated from Embryonic Stem cells.

Results: The platelets derived from embryonic stem cells are morphologically identical to normal platelets isolated from peripheral blood, and the number of viable cells was analyzed by flow cytometer. The distinctive two waves of platelet production represented normal primitive and definitive hematopoiesis.

Conclusion: This study is a preliminary milestone in platelet propagation from embryonic stem cells with C. papaya leaf extract. The differentiation of ESCs into megakaryocyte-mediated platelets with C. papaya leaf extract has shown valuable information on the positive effects that embryonic stem cells can be used for large-scale platelet production.

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References

Machlus KR, Italiano JE. The incredible journey: from megakaryocyte development to platelet formation. J Cell Biol. 2013;201(6):785-96. doi: 10.1083/jcb.201304054, PMID 23751492, PMCID PMC3678154.

Junt T, Schulze H, Chen Z, Massberg S, Goerge T, Krueger A. Dynamic visualization of thrombopoiesis within bone marrow. Science. 2007;317(5845):1767-70. doi: 10.1126/science.1146304, PMID 17885137.

Kowata S, Isogai S, Murai K, Ito S, Tohyama K, Ema M. Platelet demand modulates the type of intravascular protrusion of megakaryocytes in bone marrow. Thromb Haemost. 2014;112(4):743-56. doi: 10.1160/TH14-02-0123, PMID 24965909.

Guerriero R, Testa U, Gabbianelli M, Mattia G, Montesoro E, Macioce G. Unilineage megakaryocytic proliferation and differentiation of purified hematopoietic progenitors in serum-free liquid culture. Blood. 1995;86(10):3725-36. doi: 10.1182/blood.V86.10.3725.bloodjournal86103725, PMID 7579339.

Hagiwara T, Kodama I, Horie K, Kato T, Miyazaki H. Proliferative properties of human umbilical cord blood megakaryocyte progenitor cells to human thrombopoietin. Exp Hematol. 1998;26(3):228-35. PMID 9502619.

Choi ES, Nichol JL, Hokom MM, Hornkohl AC, Hunt P. Platelets generated in vitro from proplatelet-displaying human megakaryocytes are functional. Blood. 1995;85(2):402-13, PMID 7529062.

Bruno E, Cooper RJ, Briddell RA, Hoffman R. Further examination of the effects of recombinant cytokines on the proliferation of human megakaryocyte progenitor cells. Blood. 1991;77(11):2339-46, PMID 1710149.

Bruno E, Cooper RJ, Wilson EL, Gabrilove JL, Hoffman R. Basic fibroblast growth factor promotes the proliferation of human megakaryocyte progenitor cells. Blood. 1993;82(2):430-5, PMID 8329701.

Keller GM. In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol. 1995;7(6):862-9. doi: 10.1016/0955-0674(95)80071-9, PMID 8608017.

Wang Q, Cooney AJ. The role of nuclear receptors in embryonic stem cells. Adv Exp Med Biol. 2013;786:287-306. doi: 10.1007/978-94-007-6621-1_16, PMID 23696363.

Fujimoto TT, Kohata S, Suzuki H, Miyazaki H, Fujimura K. Production of functional platelets by differentiated embryonic stem (ES) cells in vitro. Blood. 2003;102(12):4044-51. doi: 10.1182/blood-2003-06-1773, PMID 12920021.

Morrison SJ, Uchida N, Weissman IL. The biology of hematopoietic stem cells. Annu Rev Cell Dev Biol. 1995;11:35-71. doi: 10.1146/annurev.cb.11.110195.000343. PMID 8689561.

Nakano T, Kodama H, Honjo T. Generation of lymphohematopoietic cells from embryonic stem cells in culture. Science. 1994;265(5175):1098-101. doi: 10.1126/science.8066449, PMID 8066449.

Kumar PS, Chandrasekhar C, Srikanth L, Sarma PVGK. In vitro large scale production of megakaryocytes to functional platelets from human hematopoietic stem cells. Biochem Biophys Res Commun. 2018;505(1):168-75. doi: 10.1016/j.bbrc.2018.09.090. PMID 30243726.

Sathyapalan DT, Padmanabhan A, Moni M, P-Prabhu B, Prasanna P, Balachandran S. Efficacy & safety of Carica papaya leaf extract (CPLE) in severe thrombocytopenia (≤30,000/μl) in adult dengue–results of a pilot study. PLOS ONE. 2020;15(2):e0228699. doi: 10.1371/journal.pone.0228699. PMID 32074143.

Singh SP, Kumar S, Mathan SV, Tomar MS, Singh RK, Verma PK. Therapeutic application of Carica papaya leaf extract in the management of human diseases. Daru. 2020;28(2):735-44. doi: 10.1007/s40199-020-00348-7, PMID 32367410, PMCID PMC7704890.

Sundarmurthy D, RJ, CL. Effect of carica papaya leaf extract on platelet count in chemotherapy-induced thrombocytopenic patients: a preliminary study. Natl J Physiol Pharm Pharmacol. 2017;7(6):1. doi: 10.5455/njppp.2017.7.0202628022017.

Adarsh DB, Chandrasagar K, Elango EM. Evaluation of papaya leaf extract in platelet propagation from stem cells. Int J Pharm Pharm Sci. 2023;15(2):54-9.

Weiss ML, Medicetty S, Bledsoe AR, Rachakatla RS, Choi M, Merchav S. Human umbilical cord matrix stem cells: preliminary characterization and effect of transplantation in a rodent model of Parkinson’s disease. Stem Cells. 2006;24(3):781-92. doi: 10.1634/stemcells.2005-0330, PMID 16223852.

Sarugaser R, Lickorish D, Baksh D, Hosseini MM, Davies JE. Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells. 2005;23(2):220-9. doi: 10.1634/stemcells.2004-0166, PMID 15671145.

McRedmond JP, Park SD, Reilly DF, Coppinger JA, Maguire PB, Shields DC. Integration of proteomics and genomics in platelets: a profile of platelet proteins and platelet-specific genes. Mol Cell Proteomics. 2004;3(2):133-44. doi: 10.1074/mcp.M300063-MCP200. PMID 14645502.

O’Malley CJ, Rasko JE, Basser RL, McGrath KM, Cebon J, Grigg AP. Administration of pegylated recombinant human megakaryocyte growth and development factor to humans stimulates the production of functional platelets that show no evidence of in vivo activation. Blood. 1996;88(9):3288-98. doi: 10.1182/blood.V88.9.3288.bloodjournal8893288, PMID 8896392.

Yang J, Luan J, Shen Y, Chen B. Developments in the production of platelets from stem cells (Review). Mol Med Rep. 2021;23(1). doi: 10.3892/mmr.2020.11645, PMID 33179095, PMCID PMC7673345.

Published

15-07-2023

How to Cite

D. B., A., and E. E. MURUGAIAN. “IN VITRO PROPAGATION OF PLATELETS FROM EMBRYONIC STEM CELLS WITH C. PAPAYA LEAF EXTRACT”. International Journal of Current Pharmaceutical Research, vol. 15, no. 4, July 2023, pp. 47-52, doi:10.22159/ijcpr.2023v15i4.3024.

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Original Article(s)