Summary

使用微型吸脂技术进行人体皮下脂肪组织取样

Published: September 27, 2021
doi:

Summary

手稿和相关视频展示了一种经皮活检技术,可从脐周区域获取皮下脂肪组织样本。该方法是研究脂肪组织内一系列参数(例如,基因或蛋白质表达、酶活性、脂质含量)的低风险和有效方法。

Abstract

对脂肪组织的研究有助于了解代谢和其他疾病。人体皮下脂肪组织是可及的。通过适当的培训和严格遵守无菌技术,研究人员可以在非临床环境中安全有效地获得皮下脂肪样本。在脐部外侧施用局部麻醉剂后,将连接到5或10mL注射器的14G针头通过皮肤插入皮下组织。在抽吸下,注射器以往复、切片运动移动以分离脂肪组织碎片。抽出柱塞足以确保脂肪组织碎片通过针头吸入注射器。单次活检可收集约200毫克组织。这种活检技术对参与者和研究人员来说都是非常安全的。活检后,参与者可以恢复大多数日常活动,尽管他们应该避免游泳和过度剧烈的活动48小时,以避免出血过多。参与者可以在一天内安全地进行2次活检,这意味着该技术可以应用于前后的急性干预研究。

Introduction

脂肪组织可以提供有关人类代谢功能的有用信息。人体皮下脂肪组织很容易获得。皮下脂肪组织提取技术在 80 年代中期首次被描述1;从那时起,初始方案得到了改进,以提高产量并提高研究参与者的耐受性。皮下脂肪组织可以从许多部位获得,最常见的是从臀部1 和腹部区域2获得。来自后者的样本可能更理想,因为它们在代谢疾病相关背景下提供了更有价值的信息3

使用微型吸脂方法的皮下脂肪组织活检可以在非临床环境中安全有效地进行。经过董事会认证的医生的适当培训并使用严格的无菌技术,研究人员可以常规进行这些活检,对参与者和研究人员的风险最小。活检团队必须由至少 2 人组成:进行活检的人和助手。

负责活检的人员的任务是确认参与者的身份,检查参与者是否可以安全地接受该程序(请参阅下面的协议步骤2.1-2.4),确保参与者在整个过程中感到舒适,确保在整个过程中保持无菌技术,执行程序,并为参与者提供口头和书面的善后程序。助手的作用是处理和快速处理获得的脂肪组织,以便以后分析和/或储存。助手还通过成为“非无菌手”来提供帮助,并确保参与者在整个过程中保持轻松。本视频和论文的目的是描述分步活检程序,以安全地从腹部区域获取皮下脂肪组织。

Protocol

注意:斯特灵大学NHS,侵入性或临床研究委员会批准了下面描述的活检程序。所有使用此程序的研究必须得到适当的独立伦理委员会的批准。活检者必须根据其机构的要求完成所述技术的正式培训。通常,这涉及由董事会认证的医生观察所描述的脂肪组织活检技术的演示,然后进行监督实践。一旦受训者在监督下对志愿者受试者进行了 10 次练习脂肪组织活检,他们将由董事会认证的医生进行检查…

Representative Results

所描述的脂肪组织活检程序是研究人员从人类志愿者那里获得皮下脂肪组织样本的一种有效且低风险的技术。我们使用上述程序对11名健康,体重正常的女性(年龄,27.4±3.3岁;体重指数(BMI),22.6±1.5 kg.m2)进行了39次皮下脂肪组织活检。所有参与者在禁食8-12小时后于07:00至10:00之间前往实验室。使用该脂肪组织活检程序的样品产量为192.0±97.1mg(范围= 32.8-393.6mg)(图4</…

Discussion

所描述的方案和相关视频提供了从腹部区域获得皮下脂肪组织样本的微型吸脂技术的分步概述。该研究小组在19个月内共进行了124次活检,对参与者没有不良反应。该程序是安全的,并且对参与者或活检团队的风险最小,前提是遵循所描述的安全措施。执行该程序的研究人员必须始终保持无菌技术(包括打开和分配无菌设备而不污染它们,适当地戴/摘下无菌手套,一般手部卫生)(以尽量减少参?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

提交人没有资金可以申报。

Materials

Item: 14 G needle
14 G x 3 1/8" 210 mm x 80 mm
B Braun 4665473 Per biopsy: 1
Item: 21 G needle
21 G x 1 1/2" 0.8 mm x 38 mm
Terumo AN*2138R1 Per biopsy: 2
Item: 26 G  needle
26 G x 1/2" microlance needle 0.45 mm x 13 mm
BD 303800 Per biopsy: 2
Item: 5 mL syringe
5 mL luer
DB plastipak 302187 Per biopsy: 2
Item: Adhesive wound dressing
Opsite Post-Op Dressing 9.5 x 8.5
Smith & Nephew 6600709 Per biopsy: 1
Item: Disposable sterile scalpel
Disposable Scalpel Sterile Blade no. 10
Swann Morton /0501 Per biopsy: 1
Item: Icepack
BlueDot Reusable Hot/Cold Pack 26.5 cm x 13.0 cm
NuCare F711 Per biopsy: 1
Item: Iodine based antiseptic
Videne antiseptic solution
Ecolab Videne 3030440 Per biopsy: q.s
Item: Lidocaine 2% w/o epinephrine
Lidocaine 2% injection 5 mL
B Braun 3558553 Per biopsy: 5 mL
Item: Non-sterile gloves
Starguard sensitive powder free nitrile gloves
Starguard SG-N-S Per biopsy: pair
Item: Sodium chloride 0.9%
Sodium chlride 0.9% w/v intravenous infusion BP
BBraun S8004-5384 Per biopsy: q.s.
Item: Stabilization solution*
RNAlater Stabilization Solution
ThermoFisher Scientific AM7020 Per biopsy: q.s
Item: Sterile forceps
Sterile forceps
Rocialle RML109-006 Per biopsy: 1
Item: Sterile gauze swabs
Non woven swabs sterile 7.5 x 7.5 cm
Prestige 1860 Per biopsy: 5
Item: Sterile gloves
Prestige soft vinyl sterile powder free medical gloves
Prestige S: P4301
M:P3302
L:P3301
Per biopsy: pair
Item: Sterile Microcentrifuge tubes
1.5 mL Sterile Microcentrifuge Tubes
StarLab I1415-5510 Per biopsy: q.s
Item: Sterile sheet
Paper plain white 90 x 90 cm
Rocialle RML 126-216 Per biopsy: 1
Item: Weighing boat
Diamond shape weigh boats
Heathrow Scientific HS1427C Per biopsy: 1
* denotes optional materials

References

  1. Beynen, A. C., Katan, M. B. Rapid sampling and long-term storage of subcutaneous adipose-tissue biopsies for determination of fatty acid composition. American Journal of Clinical Nutrition. 42 (2), 317-322 (1985).
  2. Moran, C. N., et al. Effects of diabetes family history and exercise training on the expression of adiponectin and leptin and their receptors. Metabolism. 60 (2), 206-214 (2011).
  3. Jialal, I., Devaraj, S. Subcutaneous adipose tissue biology in metabolic syndrome. Hormone Molecular Biology and Clinical Investigation. 33 (1), (2018).
  4. WHO Guidelines on hand hygiene in health care: a summary. World Health Organization Available from: https://www.who.int/gpsc/5may/tools/who_guidelines-handhygiene_summary.pdf (2009)
  5. Kettwich, L. G., et al. New device technologies for subcutaneous fat biopsy. Amyloid. 19 (2), 66-73 (2012).
  6. Preston, R. M. Aseptic technique: evidence-based approach for patient safety. British Journal of Nursing. 14 (10), 540-546 (2005).
  7. Handiyani, H., Meily Kurniawidjaja, L., Irawaty, D., Damayanti, R. The effective needle stick injury prevention strategies for nursing students in the clinical settings: a literature review. Enfermeria Clinica. 28, 167-171 (2018).
  8. Raggio, B. S., Barton, B. M., Kandil, E., Friedlander, P. L. Association of continued preoperative aspirin use and bleeding complications in patients undergoing thyroid surgery. JAMA Otolaryngology-Head & Neck Surgery. 144, 335 (2018).
  9. Cartwright, I. J., Pockley, A. G., Galloway, J. H., Greaves, M., Preston, F. E. The effects of dietary omega-3 polyunsaturated fatty acids on erythrocyte membrane phospholipids, erythrocyte deformability and blood viscosity in healthy volunteers. Atherosclerosis. 55 (3), 267-281 (1985).
  10. Hemmrich, K., Denecke, B., Paul, N. E., Hoffmeister, D., Pallua, N. RNA isolation from adipose tissue: an optimized procedure for high RNA yield and integrity. Laboratory Medicine. 41 (2), 104-106 (2010).
  11. Chachopoulos, V., et al. A technique for subcutaneous abdominal adipose tissue biopsy via a non-diathermy method. Journal of Visual Experiments: JoVE. (127), e55593 (2017).
  12. Civelek, M., et al. Genetic regulation of human adipose microRNA expression and its consequences for metabolic traits. Human Molecular Genetics. 22 (15), 3023-3037 (2013).
  13. Chen, Y. C., et al. Feeding influences adipose tissue responses to exercise in overweight men. American Journal of Physiology Endocrinology and Metabolism. 313 (1), 84-93 (2017).
  14. Mendham, A. E., et al. Exercise training results in depot-specific adaptations to adipose tissue mitochondrial function. Scientific Reports. 10 (1), 3785 (2020).
  15. Carswell, K. A., Lee, M. J., Fried, S. K. Culture of isolated human adipocytes and isolated adipose tissue. Methods in Molecular Biology. 806, 203-214 (2012).
  16. Arner, P., Andersson, D. P., Backdahl, J., Dahlman, I., Ryden, M. Weight gain and impaired glucose metabolism in women are predicted by inefficient subcutaneous fat cell lipolysis. Cell Metabolism. 28 (1), 45-54 (2018).
  17. Mutter, G., et al. Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays. BMC genomics. 5, 88 (2004).
  18. Coleman, S. Structural fat grafting: more than a permanent filler. Plastic and Reconstructive Surgery. 118, 108-120 (2018).
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Cite This Article
MacGregor, K. A., Rodriguez-Sanchez, N., Barwell, N. D., Gallagher, I. J., Moran, C. N., Di Virgilio, T. G. Human Subcutaneous Adipose Tissue Sampling Using a Mini-Liposuction Technique. J. Vis. Exp. (175), e62635, doi:10.3791/62635 (2021).

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