臺灣博碩士論文加值系統

本試驗旨在探討肉用家禽脛軟骨發育之差異，並建立肉用家禽軟骨細胞初代體外培養模式。試驗以白肉雞、有色肉雞、北京鴨與土番鴨之胚等作為研究脛軟骨生長發育之對象。為瞭解家禽胚階段其脛骨發育之情形，將白肉雞與有色肉雞胚於孵化第12-21天，北京鴨胚於孵化第16-29天，土番鴨於由孵化第17-32天，每天取其脛骨作組織切片染色，並測定葡萄糖胺聚醣與第二型膠原蛋白在脛骨切面中所佔之比例，藉以瞭解孵化時脛骨發育之形態。分別取孵化12天之雞胚、孵化16天之北京鴨胚以及孵化18天之土番鴨胚之軟骨細胞，建立肉用家禽軟骨細胞體外培養模式，連續培養15天。試驗期間，每三天逢機抽取軟骨細胞，分析各組之乳酸去氫酶、鹼性磷酸酶活性以及第二型膠原蛋白含量之差異。除此之外，利用Alizarin red S以及Alcian blue染色方式，觀察其鈣離子與葡萄糖胺聚醣之分泌型態，以比較各組之軟骨細胞在相同體外培養的模式下，生長表現之差異。結果顯示，脛軟骨組織中細胞外基質的分泌會隨著孵化時間與脛骨長度的增加而改變，可作為一檢視脛骨發育之參考指標。肉雞組與肉鴨組分別在孵化第14-15天及第23-24天時，脛骨長度與葡萄糖胺聚醣皆明顯增加，推測在此階段乃家禽脛骨發育之重要階段。體外培養模式中，肉鴨組可明顯觀測到葡萄糖胺聚醣與鈣離子沉積之現象，且所培養之軟骨細胞從第6天開始到12天，鹼性磷酸酶活性方面，有色肉雞組較白色肉雞組者高（P<0.05）。在細胞外基質方面，其第二型膠原蛋白之分泌量，肉鴨組皆高於肉雞組。綜合試驗結果，顯示此初代培養模式可使軟骨細胞成功分化，且在細胞外基質的表現上，與家禽胚發育階段之脛骨組織有相同趨勢。臺灣之肉雞普遍存在骨骼疾病方面的問題，使肉雞的育成率無法提昇，期本研究結果可幫助瞭解脛骨發育之機制，並比較不同肉用家禽其軟骨細胞在發育時之差異，可做為進一步探討影響肉雞軟骨發育的模式。

The purpose of the experiment was to investigate the physiological parameters of bone growth and to establish the avian tibial chondrocyte culture system. Native chicken, broiler, mule duck and Pekin duck were used as the experiment materials. In order to understand the tibial development process in the embryonic stage, type II collagen and glycosaminoglycans were assayed for studying of different sources morphology of chondrocyte in vivo. Growth plate from tibial cartilages of chicken embryos(12 days of incubation), Pekin duck embryos(16 days of incubation) and mule duck embryos(18 days of incubation) were cultured in vitro for 15 days. Alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities and type II collagen contents in chondrocytes were measured every 3 days during the culturing period. Alizarin red S was used to measure the degree of mineralization. The results showed that we could not only recognize meat-type poultry tibia growth characteristics in the embryonic post-stage, but also understand the secretion of extracellular matrix. We found that meat-type chicken group had a significant longer tibia length and the amount of glycosaminoglycan in tibia at day14-15 after incubation. We also found that in meat-type duck group at day 23-24 after incubation in vitro experiment, we found ALP activity of native chickens were higher than that of broilers from day 6 to day 12（P<0.05）after culturing. The ALP activity of Pekin ducks were higher than those of mule ducks (437.67 vs. 265.81 U/mg), and ALP activity of native chickens were higher than those of broilers (390.13 vs. 171.41 U/mg) in chondrocytes. The protein concentration of type II collagen in ducks was higher than that of chickens in chondrocytes. The results revealed that the duck chondrocyte had earlier characteristics of mineralization in embryo stage than those of chickens. The established culture system may be used for the study of factors that affect the chondrocytes growth in poultry.

項目 頁次
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表次 III
謝誌 IV
壹、 摘要………………………………………………… 1
貳、 緒言………………………………………………… 2
參、 文獻檢討…………………………………………… 4
肆、 材料與方法………………………………………… 14
伍、 結果與討論………………………………………… 21
陸、 結論………………………………………………… 49
柒、 參考文獻…………………………………………… 50
捌、 英文摘要…………………………………………… 55
玖、 附錄………………………………………………… 57

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