Name: 视频: 从小鼠泌尿生殖系统中提取前列腺:一种将雄性小鼠前列腺与其他泌尿和生殖系统器官分离的技术 - 概念
Uploaded: 2023-04-30T13:28:42.000+00:00
Duration: 1 min 51 s
Description: 1.5K 次观看. 来源:Nath， D. et al. 用于体外 3D 球状体培养模型的小鼠前列腺叶的鉴定、组织学特征和解剖。J. Vis. Exp. （2018 年） 在本视频中，我们描述了从雄性小鼠模型中解剖泌尿生殖系统或 UGS 的程序。该 UGS 在体外进一步解剖以分离前列腺并研究其在前列腺癌发展中的作用。

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All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. The Urogenital System (UGS) Dissection
NOTE: The schematic is presented in Figure 1.
<ol>
	<li>Euthanize a 3-month-old male C57BL/6 mouse using the CO2 inhalational euthanasia method or another approved technique. 
	NOTE: Mice between the ages of 3 and 12 months can be used successfully for the experiment. Older mice (&#62; 6 months) will most likely have more fat around the UGS, which will need to be cleared. Identification and separation of the lobes is often difficult in mice younger than 3 months. Other strains can be used for the experiment as well.</li>
	<li>Place the mouse on its back and secure the legs using pins so that the ventral side of the animal is exposed.</li>
	<li>Spray 70% ethanol on the mouse&#8217;s abdomen and wipe it clean. 
	NOTE: Shaving the abdomen hair before dissection is not required.</li>
	<li>Lift the skin from the abdomen, along with the muscle layer, with a pair of medium blunt forceps and make an inverted Y-shaped incision on the abdomen using scissors.
	<ol>
		<li>First, make a straight incision with a pair of sharp scissors from just above the penis to the sternum (Figure 2a).</li>
		<li>Cut from the base of the incision towards each toe, up to the thighs (Figure 2b). Fold back the skin on both sides and at the bottom to view the entire abdominal area (Figure 2c-e). 
		NOTE: The cut size varies depending on the mouse&#8217;s age and size. The size of the initial straight incision may range from 1.5 to 4 cm, depending on the size of the mouse.</li>
	</ol>
	</li>
	<li>Move over the other organs to expose the UGS. Lift and move the brown-yellow intestines (Figure 2f). Pick up the abdominal fat pads (the opaque white spongy tissue) with forceps and move them to the sides (Figure 2g). 
	NOTE: The UGS comprises the seminal vesicles, urethra, prostate, ductus deferens (vas deferens), and urinary bladder. It can be identified by the characteristic pair of opaque white semicircular arches (which are the seminal vesicles), with the fluid-filled bladder sac attached to the base. The translucent tissue located right under the seminal vesicles is the prostate.</li>
	<li>Locate the UGS, firmly hold the urinary bladder with blunt forceps, and lift the entire UGS upwards from the mouse abdomen. 
	NOTE: If the bladder is full, drain it first with a small syringe to provide a better grip with the forceps and reduce the risk of rupturing the bladder.</li>
	<li>Continuing to pull up on the bladder, slide a pair of scissors underneath the bladder and prostate all the way to the spine, and make a cut. Cut through any remaining connections to the abdominal cavity (Figure 2h and 2i). Be careful not to cut too close to the UGS to avoid accidental cutting through any prostate tissue. 
	NOTE: While making the cut under the UGS, the scissors should be inserted all the way to the back of the mouse, so that the cut snaps the vertebral column as well. This method results in severing nearly all of the connections between the UGS and the abdominal cavity in one snip, reducing the time needed to extract the tissue from the mouse.</li>
	<li>Remove the UGS and place it in 2-6 mL (enough to cover the tissue) of phosphate-buffered saline (PBS) or Dulbecco&#39;s modified Eagle medium (DMEM, high glucose) in a 6 cm Petri dish (Figure 2j and 2k) and move it to a dissection microscope (10X magnification). 
	NOTE: Change the dissecting media as required in the remaining steps.</li>
</ol>

<table><tbody><tr><td>Mouse surgical instruments&lt;br/&gt; (Mouse Dissecting kit)</td><td>World Precision Instruments</td><td>MOUSEKIT</td><td></td></tr><tr><td>Dissection microscope</td><td></td><td></td><td></td></tr><tr><td>Dissection medium (DMEM +&lt;br/&gt; 10%FBS)</td><td>Thermofisher Scientific</td><td>11965-084</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Thermofisher Scientific</td><td>10438018</td><td></td></tr></tbody></table>

extracting prostate from mouse urogenital system: a technique to separate prostate gland from other urinary and genital system organs in male mouse

Source: Nath, D. et al. <a href="https://jove.unicartagenaproxy.elogim.com/video/58397" target="_blank">Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models.</a> J. Vis. Exp. (2018)
In this video, we describe the procedure for the dissection of the urogenital system or UGS from a male mouse model. This UGS is further dissected ex vivo to isolate the prostate gland and study its role in prostate cancer development.

<img alt="Figure 1" src="/files/ftp_upload/20393/20393_Figure1.jpg" /> 
Figure 1: &#160;Schematic flowchart demonstrating dissection of the mouse urogenital system and isolation of the prostate. Flow chart for the protocol steps described in sections 1
<img alt="Figure 1" src="/files/ftp_upload/20393/20393_Figure2.jpg" /> 
Figure 2: Dissection of the mouse UGS....

Extracting Prostate from Mouse Urogenital System: A Technique to Separate Prostate Gland from Other Urinary and Genital System Organs in Male Mouse

Source: Nath, D. et al. Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models. J. ...

The urogenital system, or UGS, includes the organs of the urinary and reproductive systems located in proximity to each other. The seminal vesicles, prostate gland, testis, and ductus deferens are parts of the reproductive system, while the urinary bladder and urethra are parts of the urinary system.
To isolate the prostate gland, begin by positioning a euthanized male mouse in the supine position on a surgical bed. Incise the abdominal skin and muscle layer. Continue to cut across the midline to expose the abdominal cavity.
Now, locate the fluid-filled urinary bladder. Use forceps to pull it up and lift the entire UGS from the mouse&#39;s abdomen. Use scissors to cut the tissues underlying the UGS, till the spine, to sever any connections to the abdominal cavity. Place the harvested UGS, dorsal side up, in dissecting media and observe it under the microscope.
Next, snip the bladder from its base to expose the underlying prostate gland. Invert the UGS to locate the two tube-like ductus deferens. Cut them from the base leaving behind the prostate, seminal vesicles, and urethra. Trace both semicircular seminal vesicles and incise them at their base. The resultant tissue contains the prostate lobes wrapped around the urethra.

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1.5K 次观看. 来源:Nath， D. et al. 用于体外 3D 球状体培养模型的小鼠前列腺叶的鉴定、组织学特征和解剖。J. Vis. Exp. （2018 年） 在本视频中，我们描述了从雄性小鼠模型中解剖泌尿生殖系统或 UGS 的程序。该 UGS 在体外进一步解剖以分离前列腺并研究其在前列腺癌发展中的作用。 

视频: 从小鼠泌尿生殖系统中提取前列腺:一种将雄性小鼠前列腺与其他泌尿和生殖系统器官分离的技术 - 概念

Generation of Tumor Organoids from Genetically Engineered Mouse Models of Prostate Cancer

Methods based on homologous recombination to modify genes have significantly furthered biological research. Genetically engineered mouse models (GEMMs) are a rigorous method for studying mammalian development and disease. Our laboratory has developed several GEMMs of prostate cancer (PCa) that lack expression of one or multiple tumor suppressor genes using the site-specific Cre-loxP recombinase system and a prostate-specific promoter. In this article, we describe our method for necropsy of these PCa GEMMs, primarily focusing on dissection of mouse prostate tumors. New methods developed over the last decade have facilitated the culture of epithelial-derived cells to model organ systems in vitro in three dimensions. We also detail a 3D cell culture method to generate tumor organoids from mouse PCa GEMMs. Pre-clinical cancer research has been dominated by 2D cell culture and cell line-derived or patient-derived xenograft models. These methods lack tumor microenvironment, a limitation of using these techniques in pre-clinical studies. GEMMs are more physiologically-relevant for understanding tumorigenesis and cancer progression. Tumor organoid culture is an in vitro model system that recapitulates tumor architecture and cell lineage characteristics. In addition, 3D cell culture methods allow for growth of normal cells for comparison to tumor cell cultures, rarely possible using 2D cell culture techniques. In combination, use of GEMMs and 3D cell culture in pre-clinical studies has the potential to improve our understanding of cancer biology.&#160;

We show a method for necropsy and dissection of mouse prostate cancer models, focusing on prostate tumor dissection. A step-by-step protocol for generation of mouse prostate tumor organoids is also presented.

从前列腺癌的遗传工程小鼠模型生成肿瘤有机体

Methods based on homologous recombination to modify genes have significantly furthered biological research. Genetically engineered mouse models (GEMMs) ...

科研

JoVE Journal

遗传学

Quantification of Self-renewal in Murine Mammosphere Cultures

The mammary gland is characterized by extensive regeneration capacity, as it goes through massive hormonal changes throughout the life cycle of a female. The role of mammary stem cells (MaSCs) is widely studied both in the physiological/developmental context and with regards to breast carcinogenesis. In this aspect, ex vivo studies focused on MaSC properties are highly sought after. Mammosphere cultures represent a surrogate of organ formation and have become a valuable tool for both basic and translational research. Here, we present a detailed protocol for the generation of murine primary mammosphere cultures and the quantitation of MaSC growth properties. The protocol includes mammary gland collection and digestion, isolation of primary mammary epithelial cells (MECs), establishment of primary mammosphere cultures, serial passaging, quantitation of mammosphere growth parameters and interpretation of the results. As an example, we present the effect of low-level constitutive Myc expression on normal MECs leading to increased self-renewal and proliferation.

Here, we describe the implementation and interpretation of the results of an in vitro mammosphere self-renewal quantitative assay.

鼠乳层文化中自我更新的量化

The mammary gland is characterized by extensive regeneration capacity, as it goes through massive hormonal changes throughout the life cycle of a female. ...

发育生物学

Murine Prostate Micro-dissection and Surgical Castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

鼠前列腺微解剖和手术去势

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

Extracting Prostate from Mouse Urogenital System: A Technique to Separate Prostate Gland from Other Urinary and Genital System Organs in Male Mouse

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Extracting Prostate from Mouse Urogenital System: A Technique to Separate Prostate Gland from Other Urinary and Genital System Organs in Male Mouse