Sains Malaysiana 50(8)(2021): 2355-2365

http://doi.org/10.17576/jsm-2021-5008-18

Expired Platelet Concentrate as a Source of Human Platelet Lysate for Xenogeneic-Free Culture of Human Dermal Fibroblasts

(Platelet Pekat Tamat Tempoh sebagai Sumber Platelet Lisat Manusia untuk Pengkulturan Sel Fibroblas Kulit Manusia secara Bebas Xenogenik)

MUHAMMAD NAJIB FATHI BIN HASSAN¹, ZHENG YIE YAP¹, YEE LOONG TANG², MIN HWEI NG¹ & JIA XIAN LAW¹*

¹Centre for Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

²Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 14 May 2020/Accepted: 4 December 2020

ABSTRACT

Dermal fibroblasts have been used clinically to promote wound healing and to reduce wrinkles. Most of the time, fetal bovine serum (FBS) is used for the expansion of fibroblasts. In addition, chemically defined medium can also be used for fibroblast expansion. Nonetheless, both FBS and chemically defined medium are not ideal to culture cells that will be used clinically as FBS has the risk of pathogen transmission and induction of xenogeneic immune response whilst chemically defined medium is extremely expensive. In this study, we examine the potential of using human platelet lysate (hPL) prepared from expired platelet concentrates to culture human dermal fibroblasts. For the experiments, fibroblasts were cultured with 5 and 10% hPL, with 10% FBS as the control group to compare the cell morphology, viability, growth rate, extracellular matrix gene expression and wound healing. Results showed that fibroblasts cultured with hPL were more elongated and smaller in size. The cell viability was higher than 90% for all groups. Expansion with 10% hPL significantly shorten the population doubling time compared to the 5% hPL and 10% FBS groups. However, fibroblasts cultured with hPL have lower expression of type I collagen, type III collagen and fibronection as well as slower wound closure. In summary, hPL has the potential to be used as a serum substitute for FBS to expand fibroblasts as it significantly increases the cell proliferation. However, further studies are required to determine if the changes in the ECM gene expression and migration of the hPL-expanded fibroblasts will affect the efficacy of the cells in promoting in vivo wound healing.

Keywords: Cell culture; fibroblast; human platelet lysate; platelet concentrate; skin

ABSTRAK

Sel fibroblas kulit telah digunakan secara klinikal untuk merangsang penyembuhan luka dan mengurangkan kedut. Kebiasaanya, serum fetus lembu (FBS) paling kerap digunakan untuk pengkulturan sel fibroblas. Pada masa yang sama, media ditakrifkan secara kimia juga boleh digunakan untuk pengkulturan sel fibroblas. Namun demikian, kedua-dua FBS and media ditakrifkan secara kimia adalah tidak ideal untuk penghasilan besar-besaran sel fibroblas untuk kegunaan klinikal kerana FBS mempunyai risiko penjalaran patogen haiwan dan penolakan oleh sistem imun pesakit manakala media tanpa serum adalah sangat mahal. Dalam kajian ini, potensi platelet lisat manusia (hPL) yang disediakan daripada platelet pekat tamat tempoh untuk pengkulturan sel fibroblas telah dikaji. Untuk tujuan uji kaji, sel fibroblas dikultur dengan 5% hPL dan 10% hPL dengan 10% FBS sebagai kumpulan kawalan untuk menilai perbezaan morfologi sel, kebolehhidupan sel, kadar pertumbuhan sel, ekspresi gen-gen matriks extrasel dan penyembuhan luka. Hasil kajian mendapati sel fibroblas dikultur dengan hPL adalah lebih gelendong dan kecil berbanding dengan sel yang dikultur dengan FBS. Kebolehhidupan sel melebihi 90% untuk semua kumpulan kajian. Kadar pertumbuhan sel adalah lebih tinggi untuk kumpulan 10% hPL berbanding dengan kumpulan yang lain. Namun demikian, sel yang dikultur dengan hPL mempunyai ekspresi kolagen kumpulan I, kolagen kumpulan III dan fibronectin yang lebih rendah serta penyembuhan luka yang lebih lambat. Secara kesimpulan, hPL berpotensi menggantikan FBS untuk pengkulturan sel fibroblas kerana ia merangsang proliferasi sel. Namun demikian, penyelidikan lanjutan perlu dijalankan untuk mengenal pasti sama ada perubahan dalam ekspresi gen ekstrasel and migrasi akan menjejaskan efikasi sel-sel tersebut dalam merangsang penyembuhan luka secara in vivo.

Kata kunci: Fibroblas; kulit; pengkulturan sel; platelet lisat manusia; platelet pekat

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*Corresponding author; email: lawjx@ppukm.ukm.edu.my