我们确认梅奥诊所显微镜细胞分析核心流细胞学设施，以协助FACS分拣。

所有动物实验程序均经梅奥诊所机构动物护理和使用委员会批准。
1. 解决方案准备 
<ol><li>通过溶解拼贴酶 II 2 g 在 100 mL 汉克斯的平衡盐溶液 （HBSS） 中准备拼贴酶 2% （w/v） 溶液。每次使用均报价200μL。</li>
	<li>混合84毫升F-12介质、15毫升马血清和1毫升青霉素/链霉素，制备中和介质。</li>
	<li>通过溶解 500 毫克牛血清白蛋白 （BSA） 和 400 μL 的 0.5 M EDTA 在 100 mL 杜尔贝科的磷酸盐缓冲盐水 （DPBS） 中准备流细胞学缓冲。</li>
</ol>2. 表皮脂肪垫解剖和组织分离
<ol><li>人道安乐死（异体/颈椎错位）一只雄性小鼠。在此协议中，使用了一个四个月大的 FVB 鼠标。</li>
	<li>在腹部涂抹/喷洒 70% 乙醇，使用干净的剪刀和钳子暴露下腹腔。</li>
	<li>定位表皮脂肪垫（近似/附着在睾丸上）。用钝力握住睾丸和表皮脂肪垫之间的交界处，轻轻拉动以解放表皮脂肪垫。</li>
	<li>使用剪刀切割睾丸，在 5 mL 的 HBSS 中孵育表皮脂肪垫，在室温下在 50 mL 圆锥管中补充 3% BSA 15 分钟。</li>
	<li>离心机在150 x g 7分钟在4°C。</li>
	<li>从 50 mL 圆锥管中取出漂浮的表皮脂肪垫，用干净的剪刀细碎组织。</li>
	<li>在 13 mL 培养管中将 200μL 拼贴酶 2% 溶液添加到 3.8 mL 的 HBSS 中。加入切碎的组织，在 5 rpm 旋转孵化器中孵化，在 37 °C 下为 1 小时。 37 °C 的细胞培养孵化器可以偶尔搅拌使用，而不是旋转孵化器。</li>
	<li>将整个内容传输到 50 mL 圆锥管中，并添加 10 mL 的中和介质。通过倒置管 2-3 次轻轻混合。</li>
	<li>再准备一个装有70μm细胞过滤器的50mL圆锥管。使用细胞过滤器将消化的组织过滤到新管中。</li>
	<li>在 4 °C 下将流经传输到 15 mL 圆锥管和离心机，在 350 x g 下传输 10 分钟。</li>
	<li>小心地删除超纳坦和重新悬浮的细胞颗粒与5 mL的DPBS。离心机在350 x g 10分钟在4°C。</li>
	<li>小心地删除超纳坦，并添加 50 μL 的流细胞学缓冲区。轻轻吹笛，将细胞放在冰上，混合好。由于细胞颗粒的体积和少量的残余超常剂，流细胞学缓冲区中的细胞总体积将大于 50 μL（约 100 μL）。</li>
</ol>3. 抗体标记和荧光激活细胞分拣 （FACS）
<ol><li>将细胞与温和的管道混合后，将54μL的细胞悬架转移到1.7 mL微中心管中。加入 6 μL 的 FcR 阻塞试剂，轻轻吹笛混合。在4°C下孵化10分钟。服用 54 μL 后保存任何剩余的细胞，并将它们保存在冰中，作为第 3.5 步和第 3.6 步的未染色控制。</li>
	<li>在步骤 3.1 的孵化过程中，通过在微中心管中结合 11 μL 的抗 Sca1-APC、抗 Ter119-FITC、抗 CD31-FITC 和抗 CD45-FITC 抗体，制备抗体鸡尾酒。通过温和的管道混合好，并保持在冰上保护免受光线。</li>
	<li>用 FcR 阻断试剂孵育 10 分钟后，加入 40 μL 的抗体鸡尾酒，通过温和的管道混合。在4°C下孵化10分钟。</li>
	<li>添加 500 μL 的流细胞学缓冲区，并辅以 1 μg/mL 的 DAPI 进入染色细胞。使用带细胞滤光器扣盖的 5 mL 试管，通过温和的管道和过滤细胞很好地混合。将细胞放在冰上。</li>
	<li>在步骤 3.1 中将 500μL 的流细胞学缓冲器添加到剩余的细胞中，并通过温和的管道很好地混合。使用带细胞滤光片捕捉帽的 5 mL 试管过滤细胞。将这些细胞保持在冰上，并在下一步用作未染色的控制。</li>
	<li>将染色细胞和未染色控制带到 FACS 分析或分拣仪器中。<ol>
<li>识别和隔离 APC+/ FITC-/ DAPI-人口与图 1中显示的量子策略。这些细胞代表可行的Sca1+/Ter119-/CD31-/CD45-细胞。使用前向 （FSC） 和侧散射 （SSC） 图耗尽碎片和细胞聚合物。然后，DAPI-人口是封闭的，然后是APC+/FITC-人口的门控化。应使用未染色的控件来帮助设置界点参数。</li>
		<li>在 1.5 mL 微中心管中将细胞收集到 500μL 的流动细胞学缓冲液中。今后，所有程序都应在无菌条件下进行，以尽量减少污染。大约50，000-100，000个细胞从一只小鼠身上收集。</li>
	</ol>
</li>
	<li>使用血细胞计计算细胞以评估细胞存活率。由于细胞聚合可以干扰进一步的下游分析，也评估细胞聚合的存在，以确保这些聚合物的最低存在（+lt;5%的可行细胞数）。</li>
</ol>

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作者没有什么可透露的。

单细胞RNA测序（scRNA-seq）作为同时研究单细胞水平不同细胞群的有力工具，正迅速获得牵引力。由于样品制备成本高，吞吐量测序高，因此必须优化蜂窝输入（高可行性和纯度），以提高实验成功的可能性。一些细胞制备方案依赖于使用低旋转洗涤和基于柱的分离去除死细胞和碎片，而没有FACS分类14。然而，许多这些方法大大减少了存活的恢复细胞的数量，在许多情况下，死细?...

脂肪组织在能量代谢中起着关键作用。多余的能量以脂质的形式储存，脂肪组织能够根据营养状况和能量需求显著扩张或缩水。脂肪组织的扩张可能是由于脂肪细胞大小（肥大）和/或脂肪细胞数（增生）的增加造成的:后一过程受脂肪祖细胞1、2的增殖和分化的严格调控。在肥胖期间，脂肪组织过度扩张，组织功能障碍（包括缺氧、炎症和胰岛素抵抗）往往发展为3、4。这些并发症是许多慢性疾病的危险因素，包括高血压、糖尿病、心血管疾病、中风和癌症。因此，限制不受控制的脂肪组织扩张和缓解脂肪组织病理学是生物医学研究的首要任务。在脂肪组织扩张过程中，驻地脂肪组织衍生的干细胞（ASCs）会增殖并按顺序分化成前细胞（承诺的祖细胞），然后发育成成熟的脂肪细胞6。最近的单细胞RNA测序（scRNA-seq）研究表明，这些脂肪祖细胞（APC）种群（ASC和前细胞）表现出大量的分子和功能异质性7，8，9，10，11，12。例如，与前极细胞7相比，ASC在表现出低致病性分化能力的同时，也表现出更高的增殖和扩张能力。在ASC和前细胞群中报告了进一步的分子差异，尽管这些差异的功能相关性仍不清楚7。这些数据共同突出了脂肪祖细胞库的复杂性，并强调了开发和标准化工具以更好地了解和操纵这些关键细胞群的必要性。
该协议详细说明了高存活率Sca1+脂肪祖细胞群与小鼠表皮脂肪垫的分离，这些脂肪垫适合敏感的下游分析，包括单细胞研究（scRNA测序）和细胞培养。表皮脂肪垫的分离和分离按照先前描述的7，13进行，稍作修改，改善孤立的APC的可行性。简言之，表皮脂肪垫上的分离细胞沾染了对Sca1的抗体，Sca1是ASC和前细胞6、7和其他血统（林）标记的标记:Ter119（红细胞）、CD31（内皮细胞）和CD45（白细胞）。可行的 Sca1+/ Ter119-/CD31-/CD45-/DAPI-细胞然后通过荧光激活细胞排序 （FACS） 进行排序。重要的是，在最近的单细胞RNA测序研究中，成功分离和分析可行的Sca1+/Lin-脂肪祖细胞，确定了ASC和前细胞7中的功能异质亚群，验证了这一协议。

<table border="0" cellpadding="0" cellspacing="0" style="width:799px;" width="799">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">1.7 mL microcentrifuge tube</td>
			<td style="width:181px;">VWR</td>
			<td style="width:119px;">87003-294</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">13 mL culture tube</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">50-809-216</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">15 mL conical tube</td>
			<td style="width:181px;">Greiner Bio-one</td>
			<td style="width:119px;">188 271</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">5 mL test tube with cell strainer snap cap</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">08-771-23</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">50 mL conical tube</td>
			<td style="width:181px;">Greiner Bio-one</td>
			<td style="width:119px;">227 261</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">70 &#181;m cell strainer</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">22-363-548</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Anti-CD31-FITC antibody</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-102-519</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Anti-CD45-FITC antibody</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-102-491</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Anti-Sca1-APC antibody</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-102-833</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Anti-Ter119-FITC antibody</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-112-908</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">BSA</td>
			<td style="width:181px;">Gold Biotechnology</td>
			<td style="width:119px;">A-420-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Collagenase type II</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">17101-015</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">DAPI</td>
			<td style="width:181px;">Thermo Fisher</td>
			<td style="width:119px;">D1306</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Dulbecco&#39;s phosphate-buffered saline (DPBS)</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">14190-144</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">F-12 medium</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">11765-054</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">FcR blocking reagent</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-092-575</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Hanks&#39; balanced salt solution (HBSS)</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">14025-092</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Horse serum</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">16050-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Penicillin-streptomycin</td>
			<td style="width:181px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:332px;">Propidium iodide solution</td>
			<td style="width:181px;">Miltenyi Biotec</td>
			<td style="width:119px;">130-093-233</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of adipose progenitor cells from mouse epididymal adipose tissues

肥胖症和代谢性疾病，如糖尿病、心脏病和癌症，都与戏剧性的脂肪组织重塑有关。组织驻地脂肪祖细胞（APC）在脂肪组织平衡中起着关键作用，有助于组织病理学。单细胞分析技术（包括单细胞RNA测序和单细胞蛋白组学）的日益使用，通过允许在人口或组织范围内对单个细胞表达变化进行前所未有的解决，正在改变干细胞/祖细胞领域。在本文中，我们提供了详细的协议，以解剖小鼠表皮脂肪脂肪组织，分离单个脂肪组织衍生细胞，并执行荧光激活细胞排序（FACS），以丰富可行的Sca1+/CD31-/CD45-/Ter119- APCs。这些协议将允许调查人员准备适合下游分析的高质量APC，如单细胞RNA测序。

这个实验使用了四个月大的雄性FVB小鼠。在排除使用FSC/SSC地块的碎片和双胞胎后，对可行的细胞（DAPI- 人口）进行了封闭，随后选择了APC+/FITC- 人口（图1）。DAPI、APC 和 FITC 门是根据未染色的控制绘制的。在 图 1中显示的量化策略。
经过1小时的排序，通过流细胞学分析（图2，3）对隔离质量进...

Watch this Scientific Journal Video about Preparation of Adipose Progenitor Cells from Mouse Epididymal Adipose Tissues at JoVE.com

Preparation of Adipose Progenitor Cells from Mouse Epididymal Adipose Tissues

我们提出了一个简单的方法，利用荧光激活细胞分拣从小鼠表皮脂肪垫中分离出高度可行的青春期祖细胞。

从小鼠表皮脂肪脂肪组织中制备脂肪祖细胞

Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident ...

preparation-adipose-progenitor-cells-from-mouse-epididymal-adipose

Research

JoVE Journal

Biology

5.5K 次观看.  Mayo Clinic. 我们提出了一个简单的方法，利用荧光激活细胞分拣从小鼠表皮脂肪垫中分离出高度可行的青春期祖细胞。

视频: Preparation of Adipose Progenitor Cells from Mouse Epididymal Adipose Tissues

Manual Isolation of Adipose-derived Stem Cells from Human Lipoaspirates

In 2001, researchers at the University of California, Los Angeles, described the isolation of a new population of adult stem cells from liposuctioned adipose tissue that they initially termed Processed Lipoaspirate Cells or PLA cells. Since then, these stem cells have been renamed as Adipose-derived Stem Cells or ASCs and have gone on to become one of the most popular adult stem cells populations in the fields of stem cell research and regenerative medicine. Thousands of articles now describe the use of ASCs in a variety of regenerative animal models, including bone regeneration, peripheral nerve repair and cardiovascular engineering. Recent articles have begun to describe the myriad of uses for ASCs in the clinic. The protocol shown in this article outlines the basic procedure for manually and enzymatically isolating ASCs from large amounts of lipoaspirates obtained from cosmetic procedures. This protocol can easily be scaled up or down to accommodate the volume of lipoaspirate and can be adapted to isolate ASCs from fat tissue obtained through abdominoplasties and other similar procedures.

In 2001, researchers at UCLA described the isolation of a population of adult stem cells, termed Adipose-derived Stem Cells or ASCs, from adipose tissue. This article outlines the isolation of ASCs from lipoaspirates using a manual, enzymatic digestion protocol using collagenase.

从人类Lipoaspirates脂肪来源干细胞的手动隔离

In 2001, researchers at the University of  California, Los Angeles, described the isolation of a new population of adult  stem cells from liposuctioned ...

科研

生物学

Differentiation Capacity of Human Aortic Perivascular Adipose Progenitor Cells

Adipose tissue is a rich source of multi-potent mesenchymal stem cells (MSC) capable of differentiating into osteogenic, adipogenic, and chondrogenic lineages. Adipogenic differentiation of progenitor cells is a major mechanism driving adipose tissue expansion and dysfunction in response to obesity. Understanding changes to perivascular adipose tissue (PVAT) is thus clinically relevant in metabolic disease. However, previous studies have been predominately performed in the mouse and other animal models. This protocol uses human thoracic PVAT samples collected from patients undergoing coronary artery bypass graft surgery. Adipose tissue from the ascending aorta was collected and used for explantation of the stromal vascular fraction. We previously confirmed the presence of adipose progenitor cells in human PVAT with the capacity to differentiate into lipid-containing adipocytes. In this study, we further analyzed the differentiation potential of cells from the stromal vascular fraction, presumably containing multi-potent progenitor cells. We compared PVAT-derived cells to human bone marrow MSC for differentiation into adipogenic, osteogenic, and chondrogenic lineages. Following 14 days of differentiation, specific stains were utilized to detect lipid accumulation in adipocytes (Oil red O), calcific deposits in osteogenic cells (Alizarin Red), or glycosaminoglycans and collagen in chondrogenic cells (Masson&#8217;s Trichrome). While bone marrow MSC efficiently differentiated into all three lineages, PVAT-derived cells had adipogenic and chondrogenic potential, but lacked robust osteogenic potential.

The goal of this protocol is to test the ability of progenitor cells derived from human perivascular adipose tissue to differentiate into multiple cell lineages. Differentiation was compared to mesenchymal stem cells derived from human bone marrow, which is known to differentiate into adipocyte, osteocyte, and chondrocyte lineages.

人主动脉周脂肪祖细胞的分化能力

Adipose tissue is a rich source of multi-potent mesenchymal stem cells (MSC) capable of differentiating into osteogenic, adipogenic, and chondrogenic ...

Identification and Dissection of Diverse Mouse Adipose Depots

Adipose tissues are complex organs with a wide array of functions, including storage and mobilization of energy in response to local and global needs, uncoupling of metabolism to generate heat, and secretion of adipokines to regulate whole-body homeostasis and immune responses. Emerging research is identifying important regional differences in the developmental, molecular, and functional profiles of adipocytes located in discrete depots throughout the body. Different properties of the depots are medically relevant since metabolic diseases often demonstrate depot-specific effects. This protocol will provide investigators with a detailed anatomic atlas and dissection guide for the reproducible and accurate identification and excision of diverse mouse adipose tissues. Standardized dissection of discrete adipose depots will allow detailed comparisons of their molecular and metabolic characteristics and contributions to local and systemic pathologic states under various nutritional and environmental conditions.

Adipocytes exist in discrete depots and have diverse roles within their unique microenvironments. As regional differences in adipocyte character and function are uncovered, standardized identification and isolation of depots is crucial for advancement of the field. Herein, we present a detailed protocol for the excision of various mouse adipose depots.

各种鼠标脂肪库的识别和解剖

Adipose tissues are complex organs with a wide array of functions, including storage and mobilization of energy in response to local and global needs, ...

Isolation and Identification of Vascular Endothelial Cells from Distinct Adipose Depots for Downstream Applications

Vascular endothelial cells lining the wall of the vascular system play important roles in a variety of physiological processes, including vascular tone regulation, barrier functions, and angiogenesis. Endothelial cell dysfunction is a hallmark predictor and major driver for the progression of severe cardiovascular diseases, yet the underlying mechanisms remain poorly understood. The ability to isolate and perform analyses on endothelial cells from various vascular beds in their native form will give insight into the processes of cardiovascular disease. This protocol presents the procedure for the dissection of mouse subcutaneous and mesenteric adipose tissues, followed by isolation of their respective arterial vasculature. The isolated arteries are then digested using a specific cocktail of digestive enzymes focused on liberating functionally viable endothelial cells. The digested tissue is assessed by flow cytometry analysis using CD31+/CD45&#8722; cells as markers for positive endothelial cell identification. Cells can be sorted for immediate downstream functional assays or used to generate primary cell lines. The technique of isolating and digesting arteries from different vascular beds will provide options for researchers to evaluate freshly isolated vascular cells&#160;from arteries of interest and allow them to perform a wide range of functional tests on specific cell types.

This protocol details a method for the dissection of mouse adipose depots and the isolation and digestion of respective arteries to liberate and then identify the endothelial cell population. Freshly isolated cells used in downstream applications will advance the understanding of vascular cell biology and the mechanisms of vascular dysfunction.

从不同的脂肪库分离和鉴定血管内皮细胞，用于下游应用

Vascular endothelial cells lining the wall of the vascular system play important roles in a variety of physiological processes, including vascular tone ...

脂肪组织来源的干细胞分离和表征

Isolation and Identification of Mesenchymal Stem Cells Derived from Adipose Tissue of Sprague Dawley Rats

Adult mesenchymal cells have revolutionized molecular and cell biology in recent decades. They can differentiate into different specialized cell types, in addition to their great capacity for self-renewal, migration, and proliferation. Adipose tissue is one of the least invasive and most accessible sources of mesenchymal cells. It has also been reported to have higher yields compared to other sources, as well as superior immunomodulatory properties. Recently, different procedures for obtaining adult mesenchymal cells from different tissue sources and animal species have been published. After evaluating the criteria of some authors, we standardized a methodology that is applicable to different purposes and easily reproducible. A pool of stromal vascular fraction (SVF) from perirenal and epididymal adipose tissue allowed us to develop primary cultures with optimal morphology and functionality. The cells were observed adhered to the plastic surface for 24 h, and exhibited a fibroblast-like morphology, with prolongations and a tendency to form colonies. Flow cytometry (FC) and immunofluorescence (IF) techniques were used to assess the expression of the membrane markers CD105, CD9, CD63, CD31, and CD34. The ability of adipose-derived stem cells (ASCs) to differentiate into the adipogenic lineage was also assessed using a cocktail of factors (4 &#181;M insulin, 0.5 mM 3-methyl-iso-butyl-xanthine, and 1 &#181;M dexamethasone). After 48 h, a gradual loss of fibroblastoid morphology was observed, and at 12 days, the presence of lipid droplets positive to oil red staining was confirmed. In summary, a procedure is proposed to obtain optimal and functional ASC cultures for application in regenerative medicine.

作者聚焦:分离和鉴定源自 Sprague Dawley 大鼠脂肪组织的间充质干细胞  

This protocol describes a methodology for isolating and identifying adipose tissue-derived mesenchymal stem cells (MSCs) from Sprague Dawley rats.

斯普拉格道利大鼠脂肪组织间充质干细胞的分离和鉴定

Adult mesenchymal cells have revolutionized molecular and cell biology in recent decades. They can differentiate into different specialized cell types, in ...

培养从小鼠血管周围脂肪组织中分离的前脂肪细胞

Isolation, Culture, and Adipogenic Induction of Stromal Vascular Fraction-derived Preadipocytes from Mouse Periaortic Adipose Tissue

Perivascular adipose tissue (PVAT) is an adipose tissue depot that surrounds blood vessels and exhibits the phenotypes of white, beige, and brown adipocytes. Recent discoveries have shed light on the central role of PVAT in regulating vascular homeostasis and participating in the pathogenesis of cardiovascular diseases. A comprehensive understanding of PVAT properties and regulation is of great importance for the development of future therapies. Primary cultures of periaortic adipocytes are valuable for studying PVAT function and the crosstalk between periaortic adipocytes and vascular cells. This paper presents an economical and feasible protocol for the isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mouse periaortic adipose tissue, which can be useful for modeling adipogenesis or lipogenesis in vitro. The protocol outlines tissue processing and cell differentiation for culturing periaortic adipocytes from young mice. This protocol will provide the technological cornerstone at the bench side for the investigation of PVAT function.

作者聚焦：从小鼠血管周围脂肪组织中分离、培养和诱导 SVF 衍生的前脂肪细胞的成脂率 

Here, we describe the isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mouse periaortic adipose tissue, allowing for the study of perivascular adipose tissue function and its relationship with vascular cells.