| 摘要: | 由於胎牛血清(FBS)的生產和安全問題的爭議性，一可取代胎牛血清的方法則有其必要性。人類濃縮血小板血漿(PRP)和血小板裂解物(PL)已經被研究認為具有取代胎牛血清的能力，但往往受限於傳播感染和工業化製程上的困難。一項研究用於豬濃縮血小板血漿解決這些問題，但傳統的濃縮血小板血漿生產主要仍然限於產量及大規模生產的不適性。我們的目標則是透過優化豬濃縮血小板血漿生產配方上的改良，使其能大量製造替代胎牛血清。此外，人類濃縮血小板血漿已經被證實可透過刺激人體毛囊毛乳禿細胞(HFDPC)來改善頭髮生長；因此，我們的第二個目標是要觀察豬濃縮血小板血漿對人體毛囊毛乳禿細胞的影響和探討其作為頭髮護理產品成份促進頭髮生長的能力。
我們收集了豬抗凝全血，分離並通過兩不同的離心階段來濃縮血小板，得到大、小兩樣品。豬濃縮血小板血漿則利用膠原蛋白、凝血酶和氯化鈣的組合進行活化，從而得到三種不同的濃縮血小板血漿。透過酶聯免疫測定(ELISA)來測量其所含的生長因子(GF)和不同前處理後的穩定性，並以細胞存活分析試驗和同步聚合鏈反應技術來比較凍乾處理過的濃縮血小板血漿對HFDPC的影響。統計分析則是以FBS作為對照。
我們的研究結果證明，豬濃縮血小板血漿相較於FBS具有更高的生長因子，增加幅度為21%至400%，且對冷凍乾燥和表面活性劑處理下仍維持其穩定性，而HFDPC在冷凍乾燥後的濃縮血小板血漿培養下，相較於FBS並無明顯著差異。因而我們的研究證實豬濃縮血小板血漿作為FBS取代物及頭髮生長促進劑有其商業可行性。
Due to the controversial nature of Fetal Bovine Serum (FBS) production and safety concerns, there is a major interest in finding a viable FBS replacement. Human Platelet-Rich Plasma (PRP) and Platelet Lysate (PL) have been considered as potential FBS replacements, but there are concerns over disease transmission and commercial production is largely limited by supply. A study used porcine PRP to address these issues, but traditional PRP production is largely limited by yield, and unsuitable for large-scale production. Our aim is thus to optimize the porcine PRP production protocol, capable of manufacturing large amount of porcine PRP suitable as FBS replacement. Additionally, human PRP has been proven to improve hair growth by stimulating the Human Follicle Dermal Papilla Cells (HFDPC); as such, our secondary aim is to observe the effects of porcine PRP on HFDPC, and its potential as a hair growth-promoting active ingredient in a theoretical hair care product.
We collected anticoagulated porcine whole blood, and separated and concentrated the platelets through 2 different centrifugations, resulting in Small and Large scale preparations. The porcine PRP is then activated using a combination of collagen, thrombin, and Calcium Chloride (CaCl2), resulting in 3 distinct PRP formulations. Growth Factor (GF) levels and their stability under different treatments were measured using Enzyme-Linked Immunoassay (ELISA), and in vitro MTT assay and Real-Time PCR (QPCR) were carried out on HFDPC using original and freeze-dried PRP samples. In all cases, statistical analysis was performed using FBS as control.
Our results showed that porcine PRP has higher GF levels compared to FBS, with increases ranging from 21% to 400% and stable under freeze-drying and surfactant treatments, while HFDPC treated by original or freeze-dried PRP showed no significant difference in proliferation compared to FBS. In conclusion, our study is the first to confirm the commercial viability of porcine PRP as viable FBS replacement and hair growth promoter. |
