Utilization of Expired Platelet Concentrate for Production of Human Platelet Lysate as a Medium for T47D Cell Propagation

Diani Mentari, Relita Pebrina, Diah Nurpratami

Abstract


Background: Platelet concentrate (PC) has a short shelf life (5 days). Expired PC cannot be used for clinical purposes. PC is used for human platelet lysate (HPL) production, which was found to be more effective than FBS at increasing T47D cell proliferation. HPL production using expired PC has not been reported. This study aimed to investigate whether the use of HPL produced from expired PC (storage duration >5 days) can increase the proliferation of T47D cells in vitro.

Materials and methods: Expired PC samples with a shelf life of 7 and 11 days were used to produce HPL via freeze/thaw method. pH, total protein content, glucose and albumin levels were measured. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure proliferation rate and doubling time of HPL-treated T47D cells.

Results: After HPL production, the glucose level was influenced by the pH (p=0.003), and albumin level was influenced by total protein content (p=0.030). HPL stored for 7 and 11 days increased cell proliferation rate by 1.41 and 1.80 times higher than 10% FBS, respectively. HPL produced from expired PC did not cause morphological abnormality of the cells. In this study, the glucose levels affected cell proliferation (p=0.030). High glucose levels inhibited T47D cell proliferation.

Conclusion: Expired PC can be used as a potential material for HPL production, since HPL produced from expired PC increases cell proliferation rate and shortens cell doubling time.

Keywords: cell proliferation, human platelet lysate, platelet concentrate, thrombocyte, T47D


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Lohmann M, Walenda G, Hemeda H, Joussen S, Drescher W, Jockenhoevel S, et al. Donor age of human platelet lysate affects proliferation and differentiation of mesenchymal stem cells. PLoS One. 2012; 7(5): e37839, CrossRef.

Lubkowska A, Dolegowska B, Banfi G. Growth factor content in PRP and their applicability in medicine. Journal Biol Regul Homeost Agents. 2012; 26(2 Suppl 1): 3S-22S, article.

Wen YH, Lin WY, Lin CJ, Sun YC, Chang PY, Wang HY, et al. Sustained or higher levels of growth factors in platelet-rich plasma during 7-day storage. Clin Chim Acta. 2018; 483: 89-93, CrossRef.

Fernandez-Rebollo E, Mentrup B, Ebert R, Franzen J, Abagnale G, Sieben T, et al. Human platelet lysate versus fetal calf serum: These supplements do not select for different mesenchymal stromal cells. Sci Rep. 2017; 7(1): 5132, CrossRef.

Saury C, Lardenois A, Schleder C, Leroux I, Lieubeau B, David L, et al. Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches. Stem Cell Res Ther. 2018; 9(1): 128, CrossRef.

Doucet C, Ernou I, Zhang Y, Llense JR, Begot L, Holy X, et al. Platelet lysates promote mesenchymal stem cell expansion: A safety substitute for animal serum in cell-based therapy applications. J Cell Physiol. 2005; 205(2): 228–36, CrossRef.

Baik SY, Lim YA, Kang SJ, Ahn SH, Lee WG, Kim CH. Effects of platelet lysate preparations on the proliferation of HaCaT cells. Ann Lab Med. 2014; 34(1): 43-50, CrossRef.

Mohamed HE, Asker ME, Kotb NS, El Habab AM. Human platelet lysate efficiency, stability, and optimal heparin concentration required in culture of mammalian cells. Blood Res. 2020; 55(1): 35–43, CrossRef.

Mentari D, Pebrina R, Nurpratami D. Human platelet lysate (HPL) as an alternative media propagation of T47D cells line. Indones J Cancer Chemoprevent. 2020; 11(1): 36–45, CrossRef.

Asrianti D, Margono A, Swastiningtyas S, Asri ISR, Usman M, Yulianto I. Comparison of human platelet lysate and fetal bovine serum in culture media for human dental pulp stem cell proliferation. Int J Appl Pharm. 2019; 11(1): 157–9, CrossRef.

Menteri Kesehatan Republik Indonesia. Peraturan Menteri Kesehatan Republik Indonesia Nomor 91 Tahun 2015 Tentang Standar Pelayanan Transfusi Darah. Jakarta: Kementerian Kesehatan Republik Indonesia; 2015, article.

Dessels C, Durandt C, Pepper MS. Comparison of human platelet lysate alternatives using expired and freshly isolated platelet concentrates for adipose-derived stromal cell expansion. Platelets. 2019; 30(3): 356–67, CrossRef.

Costa EJ, Guimarães TMPD, de Almeida NC, de Toledo Vde P. Comparison of cytokine levels and metabolic parameters of stored platelet concentrates of the Fundação Hemominas, Belo Horizonte, Brazil. Rev Bras Hematol Hemoter. 2012; 34(2): 94-9, CrossRef.

Mentari D, Pebrina R, Nurpratami D. Pengaruh waktu simpan terhadap perubahan pH, kadar glukosa, laktat dehidrogenase (LDH), kalsium, mean platelet volume (MVP) sebagai indikator kualitas thrombocyte concentrate. Biomedika. 2020; 12(1): 7–15, article.

Mathai J, Resmi KR, Sulochana PV, Sathyabhama S, Baby Saritha G, Krishnan LK. Suitability of measurement of swirling as a marker of platelet shape change in concentrates stored for transfusion. Platelets. 2006; 17(6): 393–6, CrossRef.

Chung KW. Advances in understanding of the role of lipid metabolism in aging. Cells. 2021; 10(4): 880, CrossRef.

Gurina TS, Mohiuddin SS. Biochemistry, Protein Catabolism. Treasure Island: StatPearls; 2021, article.

Seidkhani-Nahal A, Allameh A, Soleimani M. Antioxidant and reactive oxygen species scavenging properties of cellular albumin in HepG2 cells is mediated by the glutathione redox system. Biotechnol Appl Biochem. 2019; 66(2): 163–71, CrossRef.

Nainggolan AL, Sudarwati S, Hilmanto D. Correlation of serum level of albumin with platelet count and platelet aggregation assessed by adenosine diphosphate agonist in children with nephrotic syndrome. Paediatrica Indonesiana. 2019; 59(1): 7–12, CrossRef.

Kholmukhamedov A, Jobe S. Platelet respiration. Blood Adv. 2019; 3(4): 599–602, CrossRef.

Weil BR, Abarbanell AM, Herrmann JL, Wang Y, Meldrum DR. High glucose concentration in cell culture medium does not acutely affect human mesenchymal stem cell growth factor production or proliferation. Am J Physiol Regul Integr Comp Physiol. 2009; 296(6): R1735–43, CrossRef.

Quagliaro L, Piconi L, Assaloni R, Martinelli L, Motz E, Ceriello A. Intermittent high glucose enhances apoptosis related to oxidative stress in human umbilical vein endothelial cells: The role of protein kinase C and NAD(P)H-oxidase activation. Diabetes. 2003; 52(11): 2795–804, CrossRef.

Francis GL. Albumin and mammalian cell culture: Implications for biotechnology applications. Cytotechnology. 2010; 62(1): 1–16, CrossRef.

Uddin S, Melnyk N, Foster DA. Albumin promotes the progression of fibroblasts through late G1 into S-phase in the absence of growth factors. Cell Cycle. 2020; 19(17): 2158–67, CrossRef.

Shanbhag S, Mohamed-Ahmed S, Lunde THF, Suliman S, Bolstad AI, Hervig T, et al. 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): 351, CrossRef.

Morgunova GV, Kolesnikov AV, Klebanov AA, Khokhlov AN. Senescence-associated β-galactosidase—A biomarker of aging, DNA damage, or cell proliferation restriction?. Moscow Univ Biol Sci Bull. 2015; 70(4): 165–7, CrossRef.

Phetfong J, Tawonsawatruk T, Seenprachawong K, Srisarin A, Isarankura-Na-Ayudhya C, Supokawej A. Re-using blood products as an alternative supplement in the optimisation of clinical-grade adipose-derived mesenchymal stem cell culture. Bone Joint Res. 2017; 6(7): 414–22, CrossRef.

Mittal K, Kaur R. Platelet storage lesion: An update. Asian J Transfus Sci. 2015; 9(1): 1–3, CrossRef.




DOI: https://doi.org/10.21705/mcbs.v6i2.254

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