Name: human subcutaneous adipose tissue sampling using a mini-liposuction technique
Uploaded: 2021-09-27T13:00:00
Description: Watch this Scientific Journal Video about Human Subcutaneous Adipose Tissue Sampling Using a Mini-Liposuction Technique at JoVE.com

The authors have no funding to declare.

NOTE: The University of Stirling NHS, Invasive, or Clinical Research Committee approved the biopsy procedure described below. All research studies using this procedure must be approved by the appropriate independent ethics committee. The biopsy taker must have completed formal training in the described technique in accordance with their institution&#39;s requirements. Typically, this involves observing a demonstration of the described adipose tissue biopsy technique by a board-certified physician, followed by supervised ...

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The described protocol and associated video provide a step-by-step overview of a mini-liposuction technique to obtain subcutaneous adipose tissue samples from the abdominal area. This research group has performed a total of 124 biopsies over the course of 19 months with no adverse effects in participants. The procedure is safe and associated with minimum risk to participants or the biopsy team, provided that the described safety measures are followed. Aseptic technique (including opening and dispensing of sterile equipme...

Adipose tissue can provide useful information on the metabolic function of humans. Human subcutaneous adipose tissue is readily accessible. A technique for subcutaneous adipose tissue extraction was first described in the mid-80s1; since then, the initial protocol has been improved to increase the yield and improve study participant tolerability. Subcutaneous adipose tissue can be obtained from numerous sites, most commonly from the glutei1 and abdominal area2. Samples from the latter may be more desirable as they provide more valuable information in metabolic disease-related contexts3.
Subcutaneous adipose tissue biopsy using the mini-liposuction method can be safely and efficiently performed in a non-clinical setting. Following appropriate training by a board-certified physician and using strict aseptic technique, researchers can routinely perform these biopsies with minimal risk to both participant and investigators. The biopsy team must consist of at least 2 individuals: the person who will perform the biopsy and an assistant.
The person responsible for the biopsy is tasked with confirming the participant&#39;s identity, checking the participant can safely undergo the procedure (see protocol steps 2.1-2.4 below), ensuring the participant is comfortable throughout the procedure, ensuring sterile technique is maintained throughout the procedure, carrying out the procedure, and providing the participant with verbal and written after-care procedures. The assistant&#39;s role is to handle and rapidly process the adipose tissue obtained for later analysis and/or storage. The assistant also helps by being the &#34;non-sterile hands&#34; and ensuring the participant is at ease throughout the procedure. The purpose of this video and paper is to describe the step-by-step biopsy procedure to safely obtain subcutaneous adipose tissue from the abdominal area.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Item: 14 G needle 
			14 G x 3 1/8&#34; 210 mm x 80 mm</td>
			<td>B Braun</td>
			<td>4665473</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: 21 G needle 
			21 G x 1 1/2&#34; 0.8 mm x 38 mm</td>
			<td>Terumo</td>
			<td>AN*2138R1</td>
			<td>Per biopsy: 2</td>
		</tr>
		<tr>
			<td>Item: 26 G&#160; needle 
			26 G x 1/2&#34; microlance needle 0.45 mm x 13 mm</td>
			<td>BD</td>
			<td>303800</td>
			<td>Per biopsy: 2</td>
		</tr>
		<tr>
			<td>Item: 5 mL syringe 
			5 mL luer</td>
			<td>DB plastipak</td>
			<td>302187</td>
			<td>Per biopsy: 2</td>
		</tr>
		<tr>
			<td>Item: Adhesive wound dressing 
			Opsite Post-Op Dressing 9.5 x 8.5</td>
			<td>Smith &#38; Nephew</td>
			<td>6600709</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: Disposable sterile scalpel 
			Disposable Scalpel Sterile Blade no. 10</td>
			<td>Swann Morton</td>
			<td>/0501</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: Icepack 
			BlueDot Reusable Hot/Cold Pack 26.5 cm x 13.0 cm</td>
			<td>NuCare</td>
			<td>F711</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: Iodine based antiseptic 
			Videne antiseptic solution</td>
			<td>Ecolab Videne</td>
			<td>3030440</td>
			<td>Per biopsy: q.s</td>
		</tr>
		<tr>
			<td>Item: Lidocaine 2% w/o epinephrine 
			Lidocaine 2% injection 5 mL</td>
			<td>B Braun</td>
			<td>3558553</td>
			<td>Per biopsy: 5 mL</td>
		</tr>
		<tr>
			<td>Item: Non-sterile gloves 
			Starguard sensitive powder free nitrile gloves</td>
			<td>Starguard</td>
			<td>SG-N-S</td>
			<td>Per biopsy: pair</td>
		</tr>
		<tr>
			<td>Item: Sodium chloride 0.9% 
			Sodium chlride 0.9% w/v intravenous infusion BP</td>
			<td>BBraun</td>
			<td>S8004-5384</td>
			<td>Per biopsy: q.s.</td>
		</tr>
		<tr>
			<td>Item: Stabilization solution* 
			RNAlater Stabilization Solution</td>
			<td>ThermoFisher Scientific</td>
			<td>AM7020</td>
			<td>Per biopsy: q.s</td>
		</tr>
		<tr>
			<td>Item: Sterile forceps 
			Sterile forceps</td>
			<td>Rocialle</td>
			<td>RML109-006</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: Sterile gauze swabs 
			Non woven swabs sterile 7.5 x 7.5 cm</td>
			<td>Prestige</td>
			<td>1860</td>
			<td>Per biopsy: 5</td>
		</tr>
		<tr>
			<td>Item: Sterile gloves 
			Prestige soft vinyl sterile powder free medical gloves</td>
			<td>Prestige</td>
			<td>S: P4301 
			M:P3302 
			L:P3301</td>
			<td>Per biopsy: pair</td>
		</tr>
		<tr>
			<td>Item: Sterile Microcentrifuge tubes 
			1.5 mL Sterile Microcentrifuge Tubes</td>
			<td>StarLab</td>
			<td>I1415-5510</td>
			<td>Per biopsy: q.s</td>
		</tr>
		<tr>
			<td>Item: Sterile sheet 
			Paper plain white 90 x 90 cm</td>
			<td>Rocialle</td>
			<td>RML 126-216</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>Item: Weighing boat 
			Diamond shape weigh boats</td>
			<td>Heathrow Scientific</td>
			<td>HS1427C</td>
			<td>Per biopsy: 1</td>
		</tr>
		<tr>
			<td>* denotes optional materials</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

human subcutaneous adipose tissue sampling using a mini-liposuction technique

Studies on adipose tissue are useful in understanding metabolic and other conditions. Human subcutaneous adipose tissue is accessible. With appropriate training and strict adherence to aseptic technique, subcutaneous adipose samples can be safely and efficiently obtained in a non-clinical setting by researchers. Following the administration of local anesthetic lateral to the umbilicus, a 14 G needle attached to a 5 or 10 mL syringe is inserted through the skin into the subcutaneous tissue. Under suction, the syringe is moved in a reciprocating, slicing motion to isolate fragments of adipose tissue. Withdrawing the plunger is enough to ensure that adipose tissue fragments are aspirated through the needle into the syringe. A single biopsy can collect about 200 mg of tissue. This biopsy technique is very safe for both participants and research staff. Following the biopsy, participants can resume most everyday activities, although they should avoid swimming and overly strenuous activities for 48 h to avoid excessive bleeding. Participants can safely undergo 2 biopsies within a single day, meaning that the technique can be applied in before-after acute intervention studies.

The described adipose tissue biopsy procedure is an efficient and low-risk technique for researchers to obtain subcutaneous adipose tissue samples from human volunteers. We performed 39 subcutaneous adipose tissue biopsies using the described procedure in 11 healthy, normal weight females (age, 27.4 &#177; 3.3 years; body mass index (BMI), 22.6 &#177; 1.5 kg.m2). All participants attended the laboratory between 07:00 and 10:00 following an 8-12 h fasting period. Sample yield using this adipose tissue biopsy pr...

Watch this Scientific Journal Video about Human Subcutaneous Adipose Tissue Sampling Using a Mini-Liposuction Technique at JoVE.com

Human Subcutaneous Adipose Tissue Sampling Using a Mini-Liposuction Technique

The manuscript and associated video demonstrate a percutaneous biopsy technique to obtain samples of subcutaneous adipose tissue from areas surrounding the umbilicus. This method is a low-risk and efficient way to investigate a range of parameters (e.g., gene or protein expression, enzyme activity, lipid content) within adipose tissue.

Studies on adipose tissue are useful in understanding metabolic and other conditions. Human subcutaneous adipose tissue is accessible. With appropriate ...

This technique is particularly well suited for any study that implements a before and after study design or wants to characterize a specific population and does not require excessive amounts of body post tissue. The main advantages are that the technique is relatively cost effective and almost pain free. You can perform multiple biopsies on the same participant on the same day.This technique can aid the understanding of adipose tissue physiology, in particular the understanding of disease and metabolic function. To begin, ensure that the laboratory has an appropriately private room with clean wipeable, non-porous surfaces, and a clean comfortable bed on which the participant can lie in a supine position. Then using a 70%ethanol spray and clean paper towels clean all required surfaces for the biopsy procedure.Next place appropriate sharps disposable bins and biohazard waist bags within easy reach of the area where the biopsy will be performed and the person taking the biopsy. Before the participant arrives in the laboratory, set up the equipment required for the procedure on a freshly cleaned general medical trolley. After obtaining written informed consent ask the participant to complete a written questionnaire to ensure they are not allergic to any materials used in this procedure.After verbally confirming the participant's identity, ensure that the participant understands the procedure to be carried out and potential secondary side effects including bruising, pain, and infection. In addition to written informed consent also take verbal consent from the participant. Next, describe the procedure to the participant emphasizing how the administration of the anesthetic and biopsy may feel.Also, ensure that the participant is comfortable with proceeding. Again, ensure that the participant has no allergies to the local anesthetic, certain metals and shellfish and that the participant is not taking any form of anticoagulant medication. Once the participant is lying in the supine position identify the biopsy site approximately five to 10 centimeters lateral to the umbilicus.Next, wash hands with soap and warm water according to standard medical guidelines. And place a sterile sheet on the clean trolley or work area taking care to only touch the outer edges of the sheet. Put on sterile surgical gloves using proper aseptic technique.Then have the assistant open the rest of the equipment such that it drops onto the prepared sterile sheet without any touching or contamination. Ensure that the assistant does not touch the items when removing them from their sterile wrappings. Next, instruct the assistant to dispense a small amount of iodine based solution on some sterile gauze.Then disinfect the skin in a spiraling motion moving outward from the proposed biopsy site. Remove excess liquid, if any by wiping with fresh sterile gauze. Verbally confirm the content and the expiry date of the local anesthetic vial with the assistant.Then instruct the assistant to hold the open vial upside down. And using a 21 gauge needle draw five milliliters of local anesthetic into a syringe. Dispose of the needle into the sharp spin and ensure that the syringe is free of air bubbles.Next, apply a 26 gauge needle to the syringe and expel any air bubbles. Then gently pinch the abdominal skin and adipose tissue moving it away from the abdominal wall and insert the needle horizontally into the subcutaneous tissue at an angle no greater than 10 degrees relative to the surface of the skin. Next, withdraw the syringe's plunger and additional 0.5 milliliters.If blood appears in the syringe, withdraw and reinsert the needle at a different angle, then raise a two to four millimeter diameter bleb to anesthetize the insertion area. Advance the needle into the subcutaneous tissue and administer one milliliter of lidocaine in a fan shaped pattern. Remove and dispose of the 26 gauge needle.Then apply a 21 gauge needle to the syringe and administer the remaining four milliliters of lidocaine in a fan shaped pattern. After five minutes using a sterile scalpel, gently prod the biopsy area to ensure the local anesthetic has taken effect and identify the boundaries of the anesthetized area. Then wait an additional minute or two and reassess.Once satisfied that the anesthesia is working, gently pinch the skin and adipose tissue and make a small one to two millimeter puncture in the skin. Next, apply a 14 gauge needle to a five or 10 milliliter syringe. Then while gently pinching the skin and adipose tissue, gently insert the needle through the puncture into the adipose tissue approximately centrally in the anesthetized area at an angle no greater than 10 degrees relative to the surface of the skin.Apply suction by withdrawing the plunger to approximately the 2.5 milliliter mark. Then collect the biopsy by moving the needle in a quick, backward and forwards motion to slice fragments of adipose tissue. After 30 seconds, twist the needle and syringe through 90 degrees and repeat this procedure to break up the fragments of adipose tissue which are then aspirated into the syringe by suction.After approximately one minute, remove the needle and empty the syringe content onto a layer of gauze covering a wane boat ensuring that the lumen of the needle is facing down to avoid potential blood spatter. Repeat the procedure of biopsy collection a maximum of three times as demonstrated, and check that the participant is content to proceed before repeating the procedure each time. While collecting the additional samples, instruct the assistant to process and prepare the previously collected samples for analysis or storage.Upon collection of satisfactory samples place one to two layers of sterile gauze over the puncture wound. Then place an ice pack and firmly apply pressure for approximately 10 minutes to induce homeostasis. Once homeostasis has occurred wipe away any iodine based solution or dried blood with sterile gauze, apply an adhesive wound dressing with an absorbent pad to the site.After checking that the participant feels well provide verbal and written instructions on biopsy site aftercare, emphasizing that the participants will likely exhibit some bruising for the next few days. Clear away used sharps and contaminated materials into designated sharps or clinical waste containers then clean all surfaces used in the biopsy procedure using 70%ethanol and place disposable and non-disposable items of bedding in appropriate clinical bags or disposal or cleaning respectively. In this study 39 subcutaneous adipose tissue biopsies were performed in 11 healthy normal weight females.Using this procedure, an average of 192 milligrams of sample was obtained. Choosing a biopsy site too close to the umbilicus can lead to unnecessarily extensive bruising. A list of complications experienced by participants following adipose tissue biopsies is summarized here.Ensure the skin is pinched at all times to avoid the needle from perforating the deeper layer of muscle or perforating through the other side of the skin. Practice makes perfect ensure that the training is performed by a trained physician before attempting it on actual participants. And when inserting the needle to break up the adipose tissue ensure that the person doing the biopsy is certain enough to break up the sample.

Research

JoVE Journal

Medicine

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