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| The Primary Cell Culture of Human Transferrin Transgenic Mouse Mammary Epithelia and the Study of the Cells Reponse to Bovine Prolactin |
| JIANG Shi-zhong1,2,3, YAN Ya-bin1,2,3, XIE Fei1,2,3, GONG Xiu-li1,2,3, HUANG Ying1,2,3, LV Bao-zhong1,2,3 |
1. Shanghai Children’s Hospital, Shanghai 200040, China;
2. Shanghai Children’s Hospital Affiliated to Shanghai Jiao Tong University, Shanghai 200040, China;
3. Shanghai Institute of Medical Genetics, Shanghai Jiao Tong University, Shanghai 200040, China |
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Abstract The aim of the construction of mammary gland bioreactor is to obtain the high-level expression of exogenous proteins in mammary gland. Prolactin plays an important role in the regulation of milk protein synthesis and secretion during mammalian lactation. The previous study demonstrated that bovine prolactin can efficiently promote the expression of exogenous gene (hTF) in milk of transgenic mice. To further explore the mechanism underlying this finding, the culture protocol were developed to cultivate transgenic mouse mammary epithelia and the biological effects of bovine prolactin on these cells were studied. Mammary tissue of mouse carrying human transferrin gene (hTF) were minced, digested with collagenase. Epithelial cells were then collected and cultured, subsequently purified by timing cells adherence to flask wall. Afterwards, purifed cells were transfected with pCMV-bPRL plasmids. The synthesis ability of the cells was tested by detecting the secretion of related proteins present in supernatant. Epithelial cells of transgenic mice carrying hTF were successfully cultured in vitro, hTF was shown to be presented in supernatants of cells. The level of hTF was elevated with the presence of bovine prolactin(1 208.55±20.07 pg/ml vs 3 161.32±71.62 pg/ml). The culture protocol could be employed in the near future experiments to investigate effects of bovine prolactin and environmental factors on the regulation of protein synthesis and secretion of mammary epithelial cells.
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Received: 26 December 2011
Published: 25 March 2012
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