Traditionally, foetal bovine serum (FBS) is used as a serum supplement for stem cell expansion in vitro. However, it is associated with xenoimmunisation and the transmission of animal pathogens, which may cause harm to stem cell recipients. As a safer alternative, human platelet lysate (HPL) has been introduced for propagating stem cells. Chondrocytes are expanded in vitro for cartilage repair via autologous chondrocyte implantation (ACI). In this study, we compare the efficacy of HPL prepared from expired platelet concentrates with that of FBS for promoting the proliferation and maintenance of the chondrogenic markers of primary human chondrocytes expanded in vitro. Chondrocytes were cultured in F12:Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% HPL, 10% HPL and 10% FBS. The cell morphology, viability and growth rate were examined from passage 1 (P1) to P3. RNA was isolated from P3 cells for quantitative polymerase chain reaction (qPCR) to determine the gene expression level of the chondrogenic, dedifferentiation and hypertrophic markers. HPL promoted chondrocyte proliferation without compromising cell viability. In addition, the chondrocytes cultured with HPL were smaller. However, HPL failed to maintain the chondrogenic markers, except SOX 9 (SRY-box transcription factor 9), which was upregulated, but not significantly. Nonetheless, HPL also suppressed the expression of type X collagen (Col X), a chondrocyte hypertrophic marker. In summary, we demonstrate the benefits of HPL supplementation in human chondrocyte culture, where it enhances cell proliferation and suppresses chondrocyte hypertrophy. In the future, HPL can be used for the large-scale expansion of chondrocytes for ACI.
ABSTRAKSecara tradisinya, serum anak lembu (FBS) digunakan untuk pengkulturan sel in vitro. Namun demikian, FBS dikaitkan dengan penolakan oleh sistem imun dan penularan patogen haiwan yang berkemungkinan membawa kemudaratan kepada penerima sel induk. Lisat platelet manusia (HPL) telah diperkenalkan sebagai alternatif yang lebih selamat untuk pengkulturan sel. Sel kondrosit telah dikultur secara in vitro untuk merawat rawan yang rosak melalui teknik implantasi kondrosit autologus (ACI). Pada kajian ini, kami membandingkan keberkesanan HPL yang disediakan daripada platelet pekat tamat tempoh dan FBS dalam merangsang proliferasi dan mengekalkan penanda kondrogenik sel kondrosit manusia yang dikulturkan secara in vitro. Sel kondrosit dikultur dalam medium F12:DMEM yang ditambah dengan 5% HPL, 10% HPL dan 10% FBS. Morfologi, kadar kehidupan dan kadar pertumbuhan sel daripada P1 hingga P3 dikenal pasti. RNA dipencil daripada sel pada P3 untuk qPCR untuk menentukan ekspresi gen penanda-penanda kondrogenik, penyahbezaan dan hipertrofi. Hasil kajian mendapati HPL merangsang proliferasi sel kondrosit tanpa memkompromikan kadar kehidupan sel. Selain itu, sel kondrosit yang dikultur dengan HPL menunjukkan saiz yang lebih kecil. Namun demikian, HPL gagal mengekalkan penanda-penanda kondrogenik kecuali SOX 9 yang meningkat secara t...