Wang Kai-Hung, Kao An-Pei, Chang Chia-Cheng, Lin Ta-Chin, Kuo Tsung-Cheng
Department of Obstetrics and Gynecology, Kuo General Hospital, Tainan, Taiwan; Center for Reproductive Medicine, Kuo General Hospital, Tainan, Taiwan; Department of Laboratory Medicine, Kuo General Hospital, Tainan, Taiwan.
Department of Research and Development, NeoAsia, Taipei, Taiwan.
Stem Cells Int. 2016;2016:4591310. doi: 10.1155/2016/4591310. Epub 2016 Oct 11.
We have previously reported the isolation and culture of a human breast epithelial cell type with stem cell characteristics (Type I HBEC) from reduction mammoplasty using the MSU-1 medium. Subsequently, we have developed several different normal human adult stem cell types from different tissues using the K-NAC medium. In this study, we determined whether this low calcium K-NAC medium with antioxidants (N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate) is a better medium to grow human breast epithelial cells. The results clearly show that the K-NAC medium is a superior medium for prolonged growth (cumulative population doubling levels ranged from 30 to 40) of normal breast epithelial cells that expressed stem cell phenotypes. The characteristics of these mammary stem cells include deficiency in gap junctional intercellular communication, expression of Oct-4, and the ability to differentiate into basal epithelial cells and to form organoid showing mammary ductal and terminal end bud-like structures. Thus, this new method of growing Type I HBECs will be very useful in future studies of mammary development, breast carcinogenesis, chemoprevention, and cancer therapy.
我们之前报道过使用MSU-1培养基从缩乳术中分离并培养出具有干细胞特征的人乳腺上皮细胞类型（I型人乳腺上皮细胞）。随后，我们使用K-NAC培养基从不同组织中培养出了几种不同的正常成人干细胞类型。在本研究中，我们确定这种添加了抗氧化剂（N-乙酰-L-半胱氨酸和L-抗坏血酸-2-磷酸）的低钙K-NAC培养基是否更适合培养人乳腺上皮细胞。结果清楚地表明，K-NAC培养基是用于正常乳腺上皮细胞长期生长（累积群体倍增水平范围为30至40）的优质培养基，这些细胞表达干细胞表型。这些乳腺干细胞的特征包括间隙连接细胞间通讯缺陷、Oct-4表达以及分化为基底上皮细胞并形成显示乳腺导管和终末芽样结构的类器官的能力。因此，这种培养I型人乳腺上皮细胞的新方法在未来乳腺发育、乳腺癌发生、化学预防和癌症治疗的研究中将非常有用。
