CHARACTERIZATION OF RAT MAMMARY EPITHELIAL-CELL SUBPOPULATIONS BY PEANUT LECTIN AND ANTI-THY-1.1 ANTIBODY AND STUDY OF FLOW-SORTED CELLS IN-VIVO
CHARACTERIZATION OF RAT MAMMARY EPITHELIAL-CELL SUBPOPULATIONS BY PEANUT LECTIN AND ANTI-THY-1.1 ANTIBODY AND STUDY OF FLOW-SORTED CELLS IN-VIVO
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DOI：
10.1006/excr.1993.1165
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发表时间：
1993-07-01
影响因子：
3.7
通讯作者：
CLIFTON, KH
中科院分区：
文献类型：
--
作者：
KIM, ND;CLIFTON, KH
建立了一种细胞分离方法，用于研究大鼠乳腺上皮细胞（RMEC）亚群在移植物和体外培养中的生长动力学。用异硫氰酸荧光素-花生凝集素和植红蛋白-抗Thy-1.1单克隆抗体对RMEC进行流式细胞术染色，从7 ~ 8周龄的F344雌性大鼠乳腺原代培养中，我们可以区分出4个细胞亚群：阴性（B-）、PNA+、Thy-1.1+和阳性（B+）。我们在含有10%胎牛血清和催乳素、17β-雌二醇、皮质醇、黄体酮和胰岛素的全激素培养基（CHM）中，对这些亚群的体外生长模式进行了1至14天的研究。随着培养时间的延长，PNA+和B-细胞的比例逐渐下降。Thy-1.1+细胞的比例随着培养时间的延长而稳步增加。有少量B+细胞。我们将RMEC移植到高泌乳素受体大鼠体内。在50%的移植部位产生至少一个肺泡单位所需的移植细胞的平均数量（ad50值）与克隆基因分数呈负相关。RMEC的AD50s如下：未分选的乳腺细胞在CHM中培养1 ~ 4天，~ 220；未分选细胞在CHM中培养7天，~ 550；从3天培养物中分选PNA+ RMEC， ~ 82；B +,∼350;B -,∼545;你的- 1.1 +,∼9372。我们得出结论，PNA+细胞亚群包括大多数克隆基因细胞。Thy-1.1+细胞表现为终末分化；它们很可能是肌上皮细胞。
A cell separation method was developed for studies of the growth kinetics of rat mammary epithelial cell (RMEC) subpopulations in grafts and in culturein vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin and phycoerythrin-anti-Thy-1.1 monoclonal antibody, we could distinguish four cell subpopulations from primary cultures of 7- to 8-week-old F344 female rat mammary glands: both negative (B-), PNA+, Thy-1.1+, and both positive (B+). We studied the growth patterns of these subpopulations invitrofor 1 to 14 days in complete hormone medium (CHM) containing 10% fetal bovine serum and prolactin, 17β-estradiol, cortisol, progesterone, and insulin. The fractions of PNA+ and B- cells steadily decreased with time in culture. The fraction of Thy-1.1+ cells steadily increased with time in culture. There were small numbers of B+ cells. We grafted RMEC in hyperprolactinemic recipient rats. The mean numbers of transplanted cells required to produce at least one alveolar unit in 50% of the graft sites (AD50values) are inversely related to the clonegenie fractions. AD50s of RMEC are as follows: unsorted mammary cells cultured in CHM for 1 to 4 days, ∼220; unsorted cells cultured in CHM for 7 days, ∼550; sorted PNA+ RMEC from outgrowths of 3-day cultures, ∼82; B+, ∼350; B-, ∼545; Thy-1.1+, ∼9372. We conclude that the PNA+ cell subpopulation includes most of the clonegenic cells. Thy-1.1+ cells appear to he terminally differentiated; they are likely to be myoepithelial cells.