應用人體造血前驅 / 幹細胞對中藥藥效及毒性反應之體外測試建立及評估

過去對於中草藥促進、恢復造血以及免疫功能的研究多以動物模式處以放射 Lethal Irradiation 的模式作為評估中草藥藥效之方法。本研究焦點放在應用人體臍帶血幹原細胞作體外培養以建立一可能代替動物及人體實驗評估中草藥藥效及毒性的方法，相較於動物及人體實驗具有經濟快速的優點。實驗結果發現 (1) PG 萃取物能使 CFU-GM 之細胞體積明顯增大，經染色後發現和活化的巨噬細胞之外觀非常相似 . (2) CSZ 複方萃取物對於 CFU-GM 有明顯促進 colony 增生之現象 (3) SD ， SM 萃取物 對 BFU-E and CFU-E 之形成有促進的效果 (4) 合成藥物 BJ-1 ， BJ-2 ， BJ-3 對 erythroid and myeloid colony 之形成有明顯的抑制而 TWD-1 只對 erythroid colony 之形成有明顯的抑制 (5) SC 萃取物在 100μg/ml 對於造血前驅 / 幹細胞無明顯的抑制而對 TF-1 Cell 之生長具有明顯的抑制。綜合以上實驗結果顯示中草藥物萃取成分刺激造血幹原細胞分化經由 colony 形態、大小及數目之改變分析可應用於評估藥效及毒性。關鍵字 : 中草藥藥效 體外集落培養分析 造血幹細胞

Evaluation of An In Vitro Colony Formation Assay for Chinese Herb Function and Toxicities

The highly proliferative potential colony - forming Assay (CFA) has been known as one of the most informative, reliable, and versatile short-term in vitro assay for examining stem/progenitor cell function. Chinese drugs working on hematopoietic system were usually examined by feeding mice with herb extract before or after irradiation damage of hematopoietic stem/progenitor cells (HS/PC) in bone marrow. In this study we use human cord blood HS/PC to conduct a human erythoid and myeloid colony assay (hCFA) for evaluation of Chinese herb function on blood renewing and immunity improving. Our results show that (1) the methanol extract of PG increases the size of the CFU-GM colonies significantly, and the cells are similar to the activated macrophage. (2) CSZ extract in 50% EtOH promotes cell proliferation of the CFU-GM. (3) The extract,SD, and ,SM, enhance both BFU-E and CFU-E formation. (4) The effects of chemical BJ-1, BJ-2, BJ-3, TWD-1 significantly inhibited the formation of BFU-E and CFU-E, however, TWD-1 inhibits the formation of BFU-E and CFU-E. (5) The extract, SC, at concentration 100μg/ml inhibits the all kind colony formation slightly ,but at the same concentration entirely inhibits the TF-1 cell proliferation. The above results suggest that the in vitro CFA method presented an advantage on economic and speed. In this study we concluded that changes in morphology, size, number of colony can be used to evaluate the function and toxicity of Chinese herb as well as the synthetic chemical drug candidates.Key Words: Chinese Herb Function,In Vitro Colony Formation Assay,Hematopoietic Stem Cell