Lipschutz J H, Foster B A, Cunha G R
Department of Medicine, University of California at San Francisco 94143-0452, USA.
Prostate. 1997 Jun 15;32(1):35-42. doi: 10.1002/(sici)1097-0045(19970615)32:1<35::aid-pros5>3.0.co;2-b.
Organ culture methods have long been used in the study of the prostate because effects of drugs and hormones can be examined in the absence of systemic effects.
Neonatal rat ventral prostates (VP) were grown on Millipore filters floating on fluid medium composed of Dulbecco's modified Eagle's medium/Ham's F-12 supplemented with insulin, transferrin, and hydrocortisone, and in the presence or absence of testosterone (T, 10(-8)M).
In the presence of T, ductal lumen formation occurred, ductal branching was extensive, and basal and luminal epithelial cells were identified by immunocytochemistry based on their distinctive cytokeratin profile. In the absence of T, ductal lumen formation did not occur, basal and luminal epithelial cells failed to differentiate, and there was a marked decrease in prostatic organ size relative to glands grown with T. Interestingly, DNA synthesis, as measured by counts per min (CPM) for 3H-thymidine incorporation, showed that DNA synthesis per microgram DNA at 7 days of organ culture was not inhibited by lack of T. Androgen receptor expression is another marker of prostatic epithelial differentiation, and it occurred in both the presence and absence of T.
Growth and differentiation of the neonatal rat prostate in vitro occur in a manner similar to that of the developing prostate in vivo, demonstrating that organ cultures of neonatal rat ventral prostates provide a faithful model for studying rat prostatic development and differentiation under serum-free conditions.
器官培养方法长期以来一直用于前列腺研究，因为可以在不产生全身效应的情况下检测药物和激素的作用。
将新生大鼠腹侧前列腺（VP）培养在漂浮于由补充了胰岛素、转铁蛋白和氢化可的松的杜氏改良伊格尔培养基/哈姆氏F-12组成的液体培养基上的密理博滤膜上，并在有或无睾酮（T，10⁻⁸M）的情况下进行培养。
在有T的情况下，导管腔形成，导管分支广泛，通过基于其独特细胞角蛋白谱的免疫细胞化学鉴定出基底上皮细胞和管腔上皮细胞。在没有T的情况下，导管腔未形成，基底上皮细胞和管腔上皮细胞未能分化，相对于在有T的情况下生长的腺体，前列腺器官大小明显减小。有趣的是，通过³H-胸腺嘧啶核苷掺入的每分钟计数（CPM）测量的DNA合成表明，器官培养7天时每微克DNA的DNA合成并未因缺乏T而受到抑制。雄激素受体表达是前列腺上皮分化的另一个标志物，在有T和无T的情况下均会出现。
新生大鼠前列腺在体外的生长和分化方式与体内发育中的前列腺相似，这表明新生大鼠腹侧前列腺的器官培养为在无血清条件下研究大鼠前列腺发育和分化提供了一个可靠的模型。
