作者没有致谢。

该议定书遵循了大学医院苏黎世人类研究伦理委员会的指导方针。
1. Neuromata 和患者选择的诊断
<ol><li>执行术前神经阻滞的近端, 以确定诊断。评估前臂中间三分之一的块, 排除 SBRN 交叉支配区的潜在诱因, 如侧前臂皮神经 (域)、骨间后神经 (针) 和尺神经背支,10。</li>
	<li>仅在减轻疼痛的患者中进行手术 (在疼痛的视觉模拟量表 (VAS) 上至少有5点)11。</li>
</ol>2. 手术准备
<ol><li>在手术前禁止病人进食6小时的食物或液体。</li>
	<li>将病人放在手术室的中央, 在一个沙滩椅位置的通用手术台上。</li>
	<li>将受影响的手臂/手放在手臂上休息。</li>
	<li>要求麻醉进行气管内麻醉 (芬太尼2µg/千克, 罗库0.6 毫克/千克, 异丙酚 150-250 毫克)。</li>
	<li>用聚维酮碘在吸脂术 (如内大腿和/或上/下腹) 和神经瘤切除部位对皮肤进行消毒。</li>
	<li>用不育的手术窗帘盖住病人, 留下暴露在抽脂部位和手术部位。</li>
</ol>3. 肿胀吸脂
<ol><li>在抽脂领域用 #11 刀片手术刀做几处直径约 3-5 毫米的小刺 (穿刺) 切口。切割到皮下层的距离。</li>
	<li>注射含有稀释利多卡因和肾上腺素 (500 毫升林格氏乳酸溶液, 20 毫升1% 利多卡因, 1 毫克肾上腺素) 的肿胀溶液, 通过14口径者 (浸润套管) 进入皮下脂肪层 (3 毫米深度)脂肪收获区。</li>
	<li>等待至少20分钟, 使肿胀的解决方案生效。</li>
	<li>钩3毫米收集套管 (外径3毫米, 长度26厘米), 以60毫升图米注射器和开始收割脂肪通过刺切口。 注: nondominant 手持续控制套管的放置和过程。</li>
	<li>拉回图米注射器的柱塞到60毫升的标记, 并放置一个蚊子钳沿撤回柱塞, 以保持负压在注射器和避免持续真空创造的主导手。</li>
	<li>在所提供的图米注射器支架中, 将图米注射器放置在垂直位置, 以获得至少220毫升的脂肪组织;这使得脂肪沉淀和分离的液体。</li>
	<li>在没有大量血液的情况下, 要特别注意获得高质量的脂肪移植。 注意: 当增强血液含量被注意时, 改变抽脂的区域。</li>
	<li>用简单的缝线缝合切口, 并按要求施加压力 (如压缩带)。</li>
	<li>将 lipoaspirate 转化为一次性无菌隔离系统。</li>
	<li>保持一个图米注射器与 lipoaspirate 在提供的立场为了得到脂肪分数为以后 lipofilling。</li>
</ol>4. 脂肪组织加工
<ol><li>关闭无菌耗材集。 注意: 隔离系统 (参见材料表) 通过执行湿测试来确认完整性。</li>
	<li>按照显示屏上的说明进行操作。 注意: 该过程主要是由软件控制的, 只需在控制面板上按下 "下一步" 或 "后退" 按钮, 就可对列出的步骤进行手动确认。lipoaspirate 被转移到组织收集罐。在组织收集罐中, 用林格氏乳酸溶液对 lipoaspirate 进行称量和自动清洗, 以去除残余的血细胞和润湿液。</li>
	<li>如果需要, 增加更多的脂肪。 注: 根据所收获的脂肪组织的重量, 隔离系统要求用户添加更多的脂肪或计算额外的酶试剂添加量。</li>
	<li>在隔离系统提示时添加所需的酶试剂量。 注意: 在酶消化过程中, 油脂和特别的分离得到了实现。因此, 组织处于摆动运动中。酶消化完成后, 特别自动转移到离心室中进行细胞浓度测定。在离心机室中, 特别被集中到一个细胞颗粒 (离心机速度大约 3000 x g)。最后, 细胞颗粒经历了一系列的冲洗与林格的乳酸溶液, 以减少残留酶水平的输出。当工艺完成时, 获得5毫升的清水 (特别)。</li>
</ol>5. 手术程序
<ol><li>要接近的神经瘤, 做一个皮肤切口在以前被诊断的神经瘤站点在前臂的远端三分之一。</li>
	<li>在外科手术双目放大镜放大镜 (2.5-3.5X) 的使用下, 继续解剖皮下组织, 并确定累及神经近端的听神经瘤。<ol>
<li>随后, 跟踪神经远端, 并确定了听神经瘤 (基于视觉检查)。</li>
	</ol>
</li>
	<li>一旦完全延长的神经瘤是明确的, 开始通过一个直接的反式神经元剪刀切除。如果没有发现任何神经瘤, 只会进行松。</li>
	<li>继续与其余的神经残肢的松, 直到3厘米近的手腕。 注意: 神经残端应位于主切口正中。</li>
	<li>将两到四小穿刺 (刺伤) 切口 (直径1毫米) 与手术刀一起通过表皮和真皮皮肤在主要方法周围。</li>
	<li>将二到四钝套管 (16 口径, 长度9厘米) 通过刺切口神经到树桩。 注意: 套管的尖端应该指向神经残端。</li>
	<li>用胶粘敷料固定套管的位置。</li>
	<li>将缝线周围的套管, 并留在原位缝合材料没有固定的节点。</li>
	<li>缝合主切口, 以防止移植的流出。 注意: 确保套管保持原位。</li>
</ol>6. 特别脂肪移植的应用
注: 经过大约1.5 小时的加工, 5 毫升的加工特别已准备好进一步使用。
<ol><li>在10毫升的交流注射器中抽取5毫升的特别。</li>
	<li>以同样的方式, 在对立的10毫升注射器中抽取2毫升的沉淀脂质分数。</li>
	<li>混合5毫升的特别与2毫升的沉淀脂质分数通过交替拉回一个注射器的柱塞和拉下来的沟通, 反对注射器的柱塞, 直到充分的混合得到。</li>
	<li>在两到四10毫升的注射器中, 将特别的脂肪移植物均分。</li>
	<li>通过将注射器连接到钝套管, 在神经残端周围均匀地分布准备好的特别-富脂接枝。 注: 在应用移植前, 确保套管在正确的位置, 并注意不要脱臼套管, 而应用脂肪移植。</li>
	<li>小心地取下套管。</li>
	<li>然后, 用柜绷带将小的穿刺切口带上。</li>
	<li>固定的手通过应用夹板在内在的加号位置 (掌 (MCP) 关节屈曲在 60-70, 间 (IP) 关节充分延长, 拇指在拳头投射, 手腕在背屈°)。如果只有松的神经被执行, 不需要夹板, 手臂和手腕包扎10天。</li>
</ol>7. 术后病人的处理
<ol><li>将病人转移至 post-anesthesia 恢复后, 并在病房内进行足够的监测。</li>
	<li>手术后10天继续固定手腕。</li>
	<li>在48小时后后进行绷带的第一次更换, 并按固定间隔继续 (即每隔一天直到出院)。</li>
	<li>切除缝合术后 12-14 天。</li>
	<li>在随访2、6、12和36月时对疼痛进行临床评估。</li>
</ol>

<ol>
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作者没有什么可透露的。

本研究的目的是有条不紊地说明的技术方面, 特别脂肪移植作为一种新的治疗症状性端 neuromata。
Neuromata 的 SBRN 与外科手术成功率只有 33%12。瓦伦et al.提出了一系列有利的结果, 早期随访使用神经脂肪移植 (框架) 没有添加特别8患者的痛苦 neuromata 的上肢5。然而, 疼痛减少在6月后疼痛复发在12月随访<sup cl...

对有症状的末端 neuromata 的管理 SBRN 仍然是挑战与许多现有的外科治疗。主要地, 神经瘤切除并且剩余的神经末梢被安置入邻居结构例如肌肉1,2, 骨头3, 或静脉4。这被假设通过模仿末端器官的气氛防止再生神经束的 out-sprouting。然而, 这些技术都没有被证明是优越的, 因为高的疼痛复发率和不足的疼痛减少往往被观察到。
最近, 脂肪移植周围剩余的神经残端产生了可喜的结果, 并主张作为一个新的治疗方案的痛苦结束 neuromata 的上肢5。除了作为机械屏障, 以防止继续运动和轻拍疼痛, 脂肪移植的有益方面是相当归因于细胞水平。人体脂肪组织包含多种细胞类型, 包括脂肪、肥大细胞、周、内皮细胞、血管平滑肌细胞、成纤维细胞和脂肪来源的干细胞 (干细胞) 以及造血干细胞6。这种异质细胞群被称为脂肪衍生再生细胞 (ADRCs) 或特别, 并可从 lipoaspirate 获得一个隔离系统 (见材料表) 和应用酶试剂 (见材料表)7。确切的行动机制还没有完全理解;然而, 特别已与改善血管生成, 免疫, 分化和分泌细胞外基质8。
特别的浓缩细胞数量增加到脂肪移植, 以减少吸收率和增加再生潜力的脂肪移植, 最近的研究显示疼痛复发后12月的脂肪移植没有添加特别5. 此外, 生物作用还通过减少残端的轴突发芽和随后的瘢痕坚持对神经本身产生有益的影响, 这可能导致神经卡压.
第一次临床研究描述特别脂肪移植作为一种新的手术选择治疗症状性端 neuromata SBRN 最近发表了10。一般来说, 该技术可以应用于任何发生在意外或医源性神经损伤后的听神经瘤。本次后续研究的目的是有条不紊地说明这一程序的技术方面。

<table border="0" cellpadding="0" cellspacing="0" width="688">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Cytori Celution System</td>
			<td style="width:71px;">Cytori</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">isolation system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Celase</td>
			<td style="width:71px;">Cytori</td>
			<td style="width:119px;"></td>
			<td>enyzme reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">liposuction system</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">tumescent solution (500 ml Lactated Ringers, 20 ml of 1% lidocaine and 1 mg epinephrine)</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">14-gauge infiltrator</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3 mm-collecting cannula (15, 26, 32 cm)</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">60 mL Tommey syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">mosquito clamp</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Toomey syringe stand</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">4 cannulas (fat injecting cannulas 1.5 mm, syringe NO.2, 90 mm)</td>
			<td style="width:71px;">Chalybs</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">10 mL syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">connecting device for syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">#11- and #15-blade scalpel</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">stevens scissors</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">pins pincette</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">bipolar electrocautery</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">suture material</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">adhesive dressing</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">binocular loupes magnifier glasses (2.5-3.5x)</td>
			<td style="width:71px;">Carl Zeiss</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:331px;">Statistical Package for Social Sciences, version 22</td>
			<td>IBM</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata

本研究的目的是有条不紊地说明和突出的关键步骤, 基质血管分数 (特别)-丰富脂肪移植作为一种新的治疗症状端 neuromata 外周感觉神经, 并在本研究, 特别是桡神经 (SBRN) 的浅支。尽管有许多现有的治疗方法, 持续的术后疼痛和常见的疼痛复发仍然是非常普遍的, 独立的程序评估。神经瘤被 microsurgically 切除相应地规范化的协议。而不是在邻近的解剖结构, 如肌肉或骨骼的再生神经残肢的迁移, 脂肪移植应用 perifocally 和作为机械屏障。为了降低脂肪的吸收率, 提高移植物的再生潜能, 将高浓度的特别集成在嫁接中。特别是从皮下脂肪分离的酶和机械 lipoaspirate 由一个特定的商业隔离系统。富含特别的脂肪移植在细胞水平上提供了机械屏障和各种生物效应, 包括改善血管生成、炎症和纤维化。在神经再生过程中, 机械和生物效应都有助于减少神经残端的无条理的轴突突起, 从而防止了痛苦的端 neuromata 的复发。

利用统计软件对数据进行分析, 并以标准偏差表示。应用科尔莫戈罗夫斯米尔诺夫试验验证了研究人群的非参数分布, 证实了负经验分布。根据研究人群的非参数性质, 采用魏氏对相关样本进行测试, 比较术前和术后结局。统计学意义被认为在 p 值小于 0.0510。
五例平均年龄为49.8 ±16.6 岁的患者包括...

Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

基质血管部分富集脂肪移植治疗症状性端 neuromata

本研究的目的是为了说明基质血管部分 (特别)-富集脂肪移植的技术过程, 作为一种新的治疗症状性端 neuromata。这种技术提供了机械屏障和生物作用的优势, 在细胞水平的加工和集中特别。

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a ...

stromal-vascular-fraction-enriched-fat-grafting-for-treatment

Research

JoVE Journal

Medicine

7.6K 次观看.  University Hospital Zurich. 本研究的目的是为了说明基质血管部分 (特别)-富集脂肪移植的技术过程, 作为一种新的治疗症状性端 neuromata。这种技术提供了机械屏障和生物作用的优势, 在细胞水平的加工和集中特别。

视频: Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of extracellular matrix (ECM) and a variety of mesenchymal cells, including fibroblasts, myofibroblasts, endothelial cells, pericytes and leukocytes 1-3.
The tumour-associated stroma is responsive to substantial paracrine signals released by neighbouring carcinoma cells. During the disease process, the stroma often becomes populated by carcinoma-associated fibroblasts (CAFs) including large numbers of myofibroblasts. These cells have previously been extracted from many different types of human carcinomas for their in vitro culture. A subpopulation of CAFs is distinguishable through their up-regulation of &alpha;-smooth muscle actin (&alpha;-SMA) expression4,5. These cells are a hallmark of 'activated fibroblasts' that share similar properties with myofibroblasts commonly observed in injured and fibrotic tissues 6. The presence of this myofibroblastic CAF subset is highly related to high-grade malignancies and associated with poor prognoses in patients.
Many laboratories, including our own, have shown that CAFs, when injected with carcinoma cells into immunodeficient mice, are capable of substantially promoting tumourigenesis 7-10. CAFs prepared from carcinoma patients, however, frequently undergo senescence during propagation in culture limiting the extensiveness of their use throughout ongoing experimentation. To overcome this difficulty, we developed a novel technique to experimentally generate immortalised human mammary CAF cell lines (exp-CAFs) from human mammary fibroblasts, using a coimplantation breast tumour xenograft model.
In order to generate exp-CAFs, parental human mammary fibroblasts, obtained from the reduction mammoplasty tissue, were first immortalised with hTERT, the catalytic subunit of the telomerase holoenzyme, and engineered to express GFP and a puromycin resistance gene. These cells were coimplanted with MCF-7 human breast carcinoma cells expressing an activated ras oncogene (MCF-7-ras cells) into a mouse xenograft. After a period of incubation in vivo, the initially injected human mammary fibroblasts were extracted from the tumour xenografts on the basis of their puromycin resistance 11.
We observed that the resident human mammary fibroblasts have differentiated, adopting a myofibroblastic phenotype and acquired tumour-promoting properties during the course of tumour progression. Importantly, these cells, defined as exp-CAFs, closely mimic the tumour-promoting myofibroblastic phenotype of CAFs isolated from breast carcinomas dissected from patients. Our tumour xenograft-derived exp-CAFs therefore provide an effective model to study the biology of CAFs in human breast carcinomas. The described protocol may also be extended for generating and characterising various CAF populations derived from other types of human carcinomas.

Experimental Generation of Carcinoma-Associated Fibroblasts CAFs from Human Mammary Fibroblasts

Carcinoma-associated fibroblasts (CAFs) rich in myofibroblasts present within the tumour stroma, play a major role in driving tumour progression. We developed a coimplantation tumour xengraft model for experimentally generating CAFs from human mammary fibroblasts. The protocol describes how to establish CAF myofibroblasts that acquire an ability to promote tumourigenesis.

从人类乳腺成纤维细胞癌相关的成纤维细胞的实验发电（CAFS）

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of ...

科研

医学

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

原位主动脉瓣移植在小鼠血管疾病的研究

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Three-dimensional Co-culture Model for Tumor-stromal Interaction

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal cells. The tumor microenvironment is comprised of a variety of cell types, such as fibroblasts, immune cells, vascular endothelial cells, pericytes and bone-marrow-derived cells, embedded in the extracellular matrix (ECM). Cancer-associated fibroblasts (CAFs) have a pro-tumorigenic role through the secretion of soluble factors, angiogenesis and ECM remodeling. The experimental models for cancer cell survival, proliferation, migration, and invasion have mostly relied on two-dimensional monocellular and monolayer tissue cultures or Boyden chamber assays. However, these experiments do not precisely reflect the physiological or pathological conditions in a diseased organ. To gain a better understanding of tumor stromal or tumor matrix interactions, multicellular and three-dimensional cultures provide more powerful tools for investigating intercellular communication and ECM-dependent modulation of cancer cell behavior. As a platform for this type of study, we present an experimental model in which cancer cells are cultured on collagen gels embedded with primary cultures of CAFs.

Here we present a protocol to co-culture in three-dimensions, which is useful for investigating multicellular interactions and extracellular matrix-dependent modulation of cancer cell behavior. In this experimental model, cancer cells are cultured on collagen gels embedded with human cancer-associated fibroblasts.

对于肿瘤基质相互作用的三维共培养模式

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal ...

Transplantation Into the Mouse Ovarian Fat Pad

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic transplantation of ovarian surface and tubal epithelia include intraperitoneal and intrabursal administration of cells. The respective limitations of these methods include poorly defined location of injected cells and limited space volume. Furthermore, they are poorly suited for long-term structural preservation of transplanted organs. To address these challenges, we have developed an alternative approach, which is based on the introduction of cells and tissue fragments into the mouse fat pad. The mouse ovarian fat pad is located in the immediate vicinity of the ovary and uterine tube (aka oviduct, fallopian tube), and provides a familiar microenvironment for cells and tissues of these organs. In our approach fluorescence-labeled mouse and human cells, and fragments of the uterine tube are engrafted by using minimally traumatic dorsal incision surgery. Transplanted cells and their outgrowths are easily located in the ovarian fat pad for over 40 days. Long-term transplantation of the entire uterine tube allows correct preservation of all principle tissue components, and does not result in adverse side effects, such as fibrosis and inflammation. Our approach should be uniquely applicable for answering important biological questions such as differentiation, regenerative and neoplastic potential of specific cell populations. Furthermore, it should be suitable for studies of microenvironmental factors in normal development and cancer.

We describe an ovarian fad pad transplantation assay suitable for studies of normal and transformed epithelia of the female reproductive tract. The mouse fat pad allows transplantation of large tissue fragments, is easily accessible for surgery and imaging, and provides the most favorable native environment for tissues of M&#252;llerian origin.

移植到小鼠卵巢脂肪垫

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

在耐药性颞叶癫痫患者卵圆孔电极记录的网络分析

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Trans-Tympanic Drug Delivery for the Treatment of Ototoxicity

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could interfere with the chemotherapeutic efficacy of the primary drugs. This is especially true for the drug cisplatin, whose anticancer actions are attenuated by antioxidants which provide adequate protection against hearing loss. Other current or potential otoprotective agents could pose a similar problem, if administered systemically. The application of various biologicals or protective agents directly to the cochlea would allow for high levels of these agents locally with limited systemic side effects. In this report, we demonstrate a trans-tympanic method of delivery of various drugs or biological reagents to the cochlea, which should enhance basic science research on the cochlea and provide a simple way of directing the use of otoprotective agents in the clinics. This report details a method of trans-tympanic drug delivery and provides examples of how this technique has been used successfully in experimental animals to treat cisplatin ototoxicity.

We present a technique for localized administration of drugs through the trans-tympanic route into the cochlea. Drug delivery through this route would not interfere with the anti-cancer efficacy of the chemotherapeutic drugs such as cisplatin.

经鼓膜药物治疗耳毒性的临床研究

The systemic administration of protective agents to treat drug-induced ototoxicity is limited by the possibility that these protective agents could ...

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

制备脂肪组织基质血管部分用于潜在软骨再生的临床协议

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem cells can be obtained from different tissues and several methods for isolation have been described. The presented method for the isolation of adipose-derived regenerative cells (ADRCs) can be used within many therapeutic areas because the method is a general procedure and, therefore, not limited to erectile dysfunction (ED) therapy. ED is a common and serious side effect to radical prostatectomy (RP) since ED often is not well treated with conventional therapy. Using ADRC&#8217;s as treatment for ED has attracted great interest due to the initial positive results after a single injection of cells into the corpora cavernosum. The method used for the isolation of ADRC&#8217;s is a simple, automated process, that is reproducible and ensures a uniform product. Furthermore, the sterility of the isolated product is ensured because the entire process takes place in a closed system. It is important to minimize the risk of contamination and infection since the stem cells are used for injection in humans. The whole procedure can be done within 2.5-3.5 hours and does not require a classified laboratory which eliminates the need for shipping tissue to an off-site. However, the procedure has some limitations since the minimum amount of drained lipoaspirate for the isolation device to function is 100 g.

Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

分离脂肪衍生的再生细胞用于治疗根治性前列腺切除术后的勃起功能障碍

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

建立干细胞治疗心肌梗塞心力衰竭的猪模型

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...