ISOLATION/CULTURE OF STEM CELLS OF PROSTATE EPITHELIUM

这项探索性拨款申请的目的是开发一种可行的方法，从成年大鼠的前列腺腹侧分离候选前列腺上皮干细胞并在体外培养它们。这一目标是基于这样的假设，即前列腺上皮是由单细胞型前体细胞(干细胞)衍生而来的。这项研究的长期目标是建立正常和转化的特定谱系的细胞系，这些细胞系来自大鼠前列腺上皮的干细胞。提出了两个特异性目标：特异性目标1.大鼠前列腺上皮干细胞的鉴定和分离。我们建议进一步建立明确的证据来证明基底细胞或基底细胞亚群是在培养中生长的前列腺上皮细胞的前体细胞。我们将考虑前列腺上皮不同细胞成分的不同沉降率、凋亡率和可能的表型标记物，对几种从上皮中分离干细胞的方法进行评估。具体目标2.干细胞的培养。在这个特定的目标中，我们将探索能够在体外培养和维持干细胞的途径，并开始定义培养条件，以产生由干细胞产生的特定谱系。

The objective of this exploratory grant application is to develop a feasible methodology of isolating candidate prostate epithelial stem cells from the adult rat ventral prostate and growing them in vitro. This objective is based on the hypothesis that the prostate epithelium is derived from a single cell- type progenitor (stem cell). The long-term goal of this research is to establish both normal and transformed cell lines of specific lineages that are derived from the stem cells of the rat prostate epithelium. Two specific aims are proposed: Specific aim 1. Identification and isolation of stem cells in the rat prostate epithelium. We propose to further establish unequivocal evidence to demonstrate that basal cells or a subpopulation of basal cells are the precursor cells of prostate epithelial cells that grow in culture. We will evaluate several isolation procedures of stem cells from the epithelium by taking into consideration the different sedimentation rates, apoptotic rates and putative phenotypic markers of the various cellular components of the prostate epithelium. Specific aim 2. Culture of stem cells. In this specific aim, we will explore avenues by which we will be able to culture and maintain stem cells in vitro as well as begin to define conditions of incubation to generate specific lineages that arise from stem cells.