The paracrine effect of bovine amniotic membrane stem cells on the mastitis treatment potential of lactating cows

• Main Authors: Ting, Wei-Je, 丁偉哲
• Other Authors: Peng, Shao-Yu
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/t64387

碩士 === 國立屏東科技大學 === 動物科學與畜產系所 === 106 === The bovine mastitis is a common disease of the Holstein farm in Taiwan. The main reason of mastitis is inflammatory reaction caused by microorganism invading the mammary gland. Producer usually used the antibiotic ointment to treat mastitis currently. But there was a problem of drug residue and drug resistance when antibiotic was abused. We would like to use the conditioned DPBS of amniotic membrane stem cells (AMSCs) to resolve this problem. We need to identify whether the cells are the stem cells before the treatment. And further to prove the differentiation ability of stem cells. AMSCs belonged to multipotent. This stem cells have lots of advantages such as noninvasive accessibility, without ethical objections and retention the immunomodulatory abilities. So there isn’t rejection or teratoma formation when the stem cells transplantation. This presentation aims to investigate the identification, differentiation abilities of AMSCs and the mastitis treatment potential with the conditioned DPBS of AMSCs. It was demonstrated that bovine AMSCs expressed CD44、CD29、CD73、CD105、CD166、Myc、Oct-4 and MHC-I. Human AMSCs can be induced to different into osteocytes, chondrocytes, adipocytes and nerve cells. On the medical application, the conditioned medium of mesenchymal stem cells contained the cytokines, hormone, peptide and mRNA secreted by stem cells. It promoted cell migration, cell proliferation and angiogenesis. So it can accelerate wound healing and suppress inflammatory reactions. Our research divided into three parts. Experiment 1 was the primary cell cultured, cell established and the identification of AMSCs. The CD44 expression was 98.6%. CD4 expressed 1.2%. It didn’t express CD105. Experiment 2 was determined the differentiation abilities of AMSCs. AMSCs could be induced to differentiate into osteocytes, chondrocytes and adipocytes. Used variety of stain to improve the successful of differentiation. Experiment 3 was injecting the conditioned DPBS of AMSCs or tetraneomycin into mastitis udder. On the ionic calcium concentration measurement of the stem cell conditioned medium treatment group, day 3 was higher than day 12 significantly. On the antibiotic control group, day 0 was higher than day 12 significantly. There was no difference between two groups. On the measurement of somatic cell counts, 60% of control group had reducing trend. 66.7% of 60% were decreased below 1 million per milliliter. At the treatment group, 80% had reducing trend. 50% of 80% were decreased below 1 million per milliliter. Above all, after identified and make sure the differentiation abilities of AMSCs. It could use the cytokines which is paracrine product of AMSCs to suppress inflammatory reactions. So it can be applied on the mastitis treatment effectively.