이 작품 (612547 및 319010 계약 번호 부여) 프로그램 유럽 연합의 지평선 2020 (부여 계약 번호 645710)와 7 프레임 워크에서 자금을 받았다. 저자 생 검 절차와 지원에 대 한 박사 경관을 Sourli-Chasioti 감사 하 고 싶습니다. 저자는 또한 비디오의 준비에서 그의 귀중 한 도움 씨 해당 Paterakis를 감사합니다.

연구 헬싱키의 선언에 의해 설정 된 표준 규격에 맞 췄 고 티 살 리의 대학, 운동 과학, Βiοethics 위원회에 의해 승인 되었다.
 1. 수술 장비 및 소모품의 준비 <ol><li>4 륜 수술 메이 요 서에 소독된 수술 필드 위치.</li>
	<li>4 륜 수술 메이 요 서에는 다음 소모품 및 수술 장비 위치: 
	바로 운영가 위 (15 cm)의 한 쌍 한 메스 (번호 11) 한 메스 핸들 곡선된가 위 (14 cm)의 한 쌍 한 모기 겸 자 Kocher 핀셋 한 켤레 한 외과 족집게 Povidone-요오드 (에 대 한 살 균 거 즈) 한 봉합 (4.0) 직선 위 (11 cm)의 한 쌍 한 바늘 홀더 (15 cm) 5 살 균 gauzes 한 접착제 멸 균 거 즈 소독된 바늘으로 한 10 mL 주사기 2 %xylocaine (아드레날린)의 10 mL 하나 소독된 외과 장갑의 쌍</li> <li>별도 벤치에 다음 자료 위치: 두 microcentrifuge 튜브 10% 포 르 말린의 5 mL를 포함 하는 1 개의 관 -190 ° c.에 액체 질소의 50-100 mL를 포함 하는 하나의 작은 컨테이너 </li></ol> 2. 참가자의 준비 <ol><li>참가자 따라 생 검 절차 전에는 8 h 빠른 15.</li>
	<li>있고 참가자 자제 운동, 과도 한 스트레스, 알코올, 그리고 액티브 간접 흡연 72 h 이전 생 16 , 17.</li>
</ol> 3. 피하 지방 조직 생 검을 수행 <ol><li>부정사 위치에 수술 침대에 참가자를 위치.</li>
	<li>절 개 povidone-요오드를 사용 하 여 영역을 소독.</li>
	<li>천천히 그리고 꾸준히 2 %xylocaine (아드레날린)의 10 mL를 주사 취 역할을 선택한 지역에 18.</li>
	<li>절 개 영역을 제외한 참가자의 시체를 충당 하기 위해 지역 무 균 수술 필드 위치.</li>
	<li>적어도 3-5 분 나중에, 로컬 마 취의 영역에 핀셋의 쌍을 가진 압력을 적용 하 고 그 느낌 그 취를 달성을 보장 하기 위해 모든 고통을 19 참가자에 게 물어.</li>
	<li>11 호 메스 메스 핸들 연결.</li>
	<li>배꼽의 오른쪽 또는 왼쪽에 3-5 cm, 길이 2-2.5 cm의 절 개를 만듭니다.</li>
	<li>스트레이트 15 cm가 위를 운영 한 켤레와 피하 조직을 제거.</li>
	<li>지방 조직 밝혀 일단 한 쌍의 족집게와 곡선된 14 cm가 포착.</li>
	<li>잘라 약 1g의 지방 조직 제거.</li>
	<li>조직 제거, 다음 놓고 살 균 gauzes hemostasis 달성 하기 위해 절 개.</li>
	<li>Hemostasis 확인 될 때 캡처 14 cm 바늘 4.0 봉합 및 피부 외과 핀셋의 쌍을 가진 피하 조직 표시 될 때까지.</li>
	<li>피하 조직으로 이동 하는 피부 및 피하는 밀접 하 게 정렬 하는 동안 피부에 봉합 바늘을 삽입.</li>
	<li>봉합 바늘, 피하 조직까지 피부 외부 구동으로 연속 지그재그 기법을 따라. 피부 절 개를 닫힐 때까지 계속.</li>
	<li>외부 봉합 넥타이.</li>
	<li>절 개 식 염 수의 영역을 청소 하 고 접착제 메 마른 거 즈로 그것을 커버.</li>
</ol> 4. 조직의 <ol><li>3 개의 조각으로 수집 된 지방 조직을 구분: 150, 350, 및 500 mg.</li>
	<li>Microcentrifuge 튜브 샘플 조직 150와 350 mg 조각 위치.</li>
	<li>-80에서 냉장고에 최종 증 착까지-190 ° C에 액체 질소로 작은 용기에 담가 두 microcentrifuge 튜브 ° C. 
	참고: 사전에 사용 하 여 무 균 바늘-190에서 액체 질소에 microcentrifuge 튜브의 침수 동안 컵 오류를 방지 하려면 microcentrifuge 튜브에 작은 구멍을 찌를 ° c.</li>
	<li>담가 샘플 조직 500mg 조각-조직학 분석을 위해 사용 될-10% 포 르 말린의 5 mL를 포함 하는 튜브에. 
	주: 수술 중 과도 한 출혈의 경우에만 모기 겸 자 사용. 이러한 경우 하 선 고 hemostasis 달성 될 때까지 닫습니다 모기 집게와 선박 포착.</li>
</ol>

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저자 들은 아무 경쟁 금융 관심사 선언 합니다.

이 연구의 목적은 복 부 피하 지방 조직 생 검, 건강 한 남성에서 비 투 열 방법으로 국 소 마 취 아래 수행에 대 한 수술 기법을 표준화 했다. 투 열은 일반적으로 수술 실;에 사용 그러나, 가능한 부작용 (즉, 플래시 화재 및 환경 독성)9 와10 치유 지연이 발생할 수 있습니다. 투 열 요법의 이러한 원치 않는 효과 운영 직원과 환자/참가자에 대 한 건강 위험?...

지방 조직 생 검 조직 샘플을 분석, 시 대사 및 질병 메커니즘의 통찰력이 개요를 제공할 수 있습니다 제공 합니다. 예를 들어, 그들은 유 방 암 종양 식별1, 지방산 조성 시험2및 중요 한 대사와 호르몬 질병 경로3에 대 한 연구에 사용 됩니다. 복 부 지역에 피하 지방 조직 생 검, 연구자와 의사는 수술 또는 바늘 기반 기술을 사용 합니다. 그러나, 수술 피하 지방 생 검 조직 샘플 [예를 들어, 메신저 ribonucleic 산 (mRNA) 식, 조직학 분석, 등] 복잡 한 생물 학적 메커니즘의 포괄적인 개요를 제공할 수 있습니다 제공할 수 있는4 . 그들은 건강 한 성인4 에서 이전에 사용 하 고 침대5에서 안전 하 게 수행할 수 있습니다.
생 검 바늘 기반 기술 조직의 무결성을 파괴 수 있습니다 강렬한 지방 흡입 필요 하 고 일반적으로 적은 양의 조직 (100-500mg)6을 검색 합니다. 이러한 조직 샘플 복잡 한 조사 연구에 적합 하지 않을 수 있습니다. 또한, 바늘 기반 기술에 비해, 수술 피하 지방 biopsies 제공할 수 있습니다 (예를 들어, B와 T 세포 신호); 여러 염증 성 지표의 더 완전 한 그림을 그릴 수 있습니다 조직 샘플 골격 규정; 세포질 통로; 지질, 탄수화물, 아미노산;의 대사 경로 그리고 산화 인 산화 통로4. 바늘-기반 기술 조직4,7,8 거리 영역에 대 한 약간의 정보를 제공 하면서 수술 한4, 보다 혈액 세포 오염에 대 한 높은 위험을 포함 하는 또한, .
일반적으로, 수술 지방 조직 생 검 투 열 치료9과도 한 출혈 방지 하기 위해 혈관을 cauterizing에 대 한 포함 되어 있습니다. 투 열 또한 구울 조직 신생 및 사마귀 질환9에 의해 감염 되는 조직에 의해 영향을 파괴 하는 데 사용 됩니다. 이 기술은 특히 신경외과 눈 수술9에 적용 됩니다.
투 열 요법의 과도 한 사용은 괴 사 성 조직, 감염을 일으키는 원인이 되 고 치유 과정10지연의 형성으로 이어질 수 있습니다. 실제로, 플래시 화재 등 조직 병 변 피부 부작용 투 열9를 다음 보고 되었습니다. 또한, 우려 참가자/환자와 운영 직원9에 의해 흡입 수 있습니다 수술 환경에 투 열에 의해 발표 외과 연기의 독성에 대해 제기 되고있다. 실제로, 투 열 요법의 사용 흡연 6 담배11 에 의해 생산에 동일 수술 실에서 어 독성을 일으킬 수 있습니다 그리고 비록 수술 일반적으로 좋은 환기 시스템, 오염 수 마지막 적어도 20 분12 . 그러나, 환자는 편도 선 수술 수술 사업, 투 열 메서드는 표시13 비-중요 한, 수술 후 통증 및 출혈14 클래식 결 찰 hemostasis 방법 보다 덜. 전반적으로, 수술 후 통증에 차이 및 건강 한 개인에서 지방 조직 생 검 후 요금을 거의 출혈 증거 존재 합니다. 투 열 요법의 부작용 및 증거의 부족을 감안할 때, 지방 조직 생 검에 대 한 비 투 열 방법의 표준화가 보증 된다. 따라서,이 연구의 목표는 복 부 피하 지방 조직 생 검, 건강 한 남자의 비 투 열 메서드를 사용 하는 국 소 마 취 아래 수행에 대 한 수술 기법을 표준화 했다. 또한, 우리는 조직 컬렉션 mRNA 표정 및 단백질 농도 결정 하 고 조직학 분석을 수행 하는 절차를 설명 합니다.

<table><tbody><tr><td>&amp;nbsp;Disinfected surgical field&amp;nbsp;</td><td>Hartmann</td><td>23516-00</td><td></td></tr><tr><td>Stainless Steel Instrument Stands (surgical Mayo)</td><td>Medical Device Depot Inc.</td><td>32000</td><td></td></tr><tr><td>Operating Scissors straight 15 cm</td><td>Karl Storz</td><td>791903</td><td></td></tr><tr><td>Scalpel No 11&amp;nbsp;</td><td>Swann-Morton</td><td>203</td><td>Bipolar, catalog number 429011</td></tr><tr><td>Scarpel handle No 3</td><td>Karl Storz</td><td>488090</td><td></td></tr><tr><td>Scissors curved 14 cm</td><td>Karl Storz</td><td>511514, 752918</td><td></td></tr><tr><td>Mosquito forceps</td><td>Karl Storz</td><td>535012</td><td></td></tr><tr><td>Tweezers Kocher&amp;nbsp;</td><td>Karl Storz</td><td>530416</td><td>Bipolar, catalog number 830316</td></tr><tr><td>Surgical tweezers</td><td>Karl Storz</td><td>793216</td><td>Bipolar, catalog number 831016</td></tr><tr><td>Povidone-iodine&amp;nbsp;</td><td>Various</td><td>Not applicable</td><td></td></tr><tr><td>Suture 4.0 Vicryl</td><td>Johnsons</td><td>V4970H&amp;nbsp;</td><td></td></tr><tr><td>Scissors straight 11 cm</td><td>Karl Storz</td><td>512511 DS</td><td></td></tr><tr><td>Needle holder 15 cm</td><td>Karl Storz</td><td>515515</td><td>Bipolar, catalog number 213015</td></tr><tr><td>&amp;nbsp;Sterile gauzes</td><td>Hartmann</td><td>3021</td><td></td></tr><tr><td>&amp;Alpha;dhesive sterile gauze&amp;nbsp;</td><td>Digas medical equipment</td><td>2889</td><td></td></tr><tr><td>10 mL syringe with disinfected needle</td><td>Alpha medical solutions</td><td>10310-55-0010</td><td></td></tr><tr><td>2%-xylocaine (no adrenaline)</td><td>Various</td><td>Not applicable</td><td></td></tr><tr><td>Sterile surgical gloves&amp;nbsp;</td><td>Sempermed</td><td>10001</td><td></td></tr><tr><td>Eppendorf tubes</td><td>Kartell</td><td>16283</td><td></td></tr><tr><td>10% formalin&amp;nbsp;</td><td>Alpha medical solutions</td><td>11016-30-2909</td><td></td></tr><tr><td>Tubes 120 mL</td><td>Digas medical equipment</td><td>9025</td><td></td></tr><tr><td>Liquid nitrogen at -190&amp;deg;C</td><td>Revival</td><td>Not applicable</td><td></td></tr></tbody></table>

a technique for subcutaneous abdominal adipose tissue biopsy via a non-diathermy method

지방 조직 생 검 조직 샘플을 분석, 시 대사 및 질병 메커니즘의 통찰력이 개요를 제공할 수 있습니다 제공 합니다. 복 부 지역에 피하 지방 조직 생 검, 연구자와 의사는 수술 또는 바늘 기반 기술을 사용 합니다. 그러나, 수술 피하 지방 biopsies 백색 지방 조직에 생물학적 지표의 복합성의 더 포괄적인 개요를 제공할 수 있는 조직 샘플을 제공할 수 있습니다. 일반적으로, 수술 지방 조직 생 검 투 열 치료 cauterizing 혈관 과도 한 출혈을 방지 하기 위해 포함 되어 있습니다. 그럼에도 불구 하 고, 플래시 화재와 조직, 피부 장애 등의 부작용 투 열 후 보고 되었습니다. 따라서, 우리는 표준화 비 투 열 방법을 사용 하 여 로컬 마 취 아래 수행 수술 복 부 지방 조직 생 검 목적입니다. 우리는 건강 한 남성 비 투 열 복 부 수술 생 검 메서드를 사용 하 여 115 피하 지방 조직 생 검 실시. 우리의 결과 115 작업 (2.61%)에서 출혈 과도 한 사후 작업의 세 가지 경우를 보여주었다. 결론적으로, 우리의 표준된 피하 복 부 지방 조직 수술 생 검 투 비 열 방법을 사용 하 여 최소한의 부작용으로 머리 맡에 건강 한 사람에 게 안전 하 게 적용할 수 있습니다.

참가자의 특성은 표 1에 제공 됩니다. 스티치 절 개의 영역에서 다음 8-12 일 이내 떨어졌다. 우리는 건강 한 성인 남성 비 투 열 복 부 수술 생 검 방법을 사용 하 여 115 피하 지방 조직 biopsies의 총 완료 (나이: 35.2 ± 6.9 년; 신체 질량 지 수: 27.3 ± 4.4; 및 체 지방 비율: 27.0 ± 9.0%). 세 가지 경우에서 관찰 되었다 과도 한 출혈 후 생 검 (보급: 2.61%)는 수술 후 2 일. 3 ?...

Watch this Scientific Journal Video about A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method at JoVE.com

A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

우리는 복 부 지방 조직 생 검 국 소 마 취 아래 수행 비 투 열 방법을 사용 하 여 표준화. 세 가지 경우의 과도 한 후 115 작업 (2.61%)에서 출혈 발생 했습니다. 우리는 복 부 지방 조직 수술 생 검 투 비 열 방법을 사용 하 여 적용할 수 있는 안전 하 게 건강 한 사람에 게 결론.

비 투 열 방법을 통해 복 부 피하 지방 조직 생 검에 대 한 기술

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To ...

a-technique-for-subcutaneous-abdominal-adipose-tissue-biopsy-via-non

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

15.0K 조회수.  University of Thessaly. 우리는 복 부 지방 조직 생 검 국 소 마 취 아래 수행 비 투 열 방법을 사용 하 여 표준화. 세 가지 경우의 과도 한 후 115 작업 (2.61%)에서 출혈 발생 했습니다. 우리는 복 부 지방 조직 수술 생 검 투 비 열 방법을 사용 하 여 적용할 수 있는 안전 하 게 건강 한 사람에 게 결론.

비디오: A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

Performing and Processing FNA of Anterior Fat Pad for Amyloid

Historically, heart, liver, and kidney biopsies were performed to demonstrate amyloid deposits in amyloidosis. Since the clinical presentation of this disease is so variable and non-specific, the associated risks of these biopsies are too great for the diagnostic yield. Other sites that have a lower biopsy risk, such as skin or gingival, are also relatively invasive and expensive. In addition, these biopsies may not always have sufficient amyloid deposits to establish a diagnosis. Fat pad aspiration has demonstrated good clinical correlation with low cost and minimal morbidity. However, there are no standardized protocols for performing this procedure or processing the aspirated specimen, which leads to variable and nonreproducible results. The most frequently utilized modality for detecting amyloid in tissue is an apple-green birefringence on Congo red stained sections using a polarizing microscope. This technique requires cell block preparation of aspirated material. Unfortunately, patients presenting in early stage of amyloidosis have minimal amounts of amyloid which greatly reduces the sensitivity of Congo red stained cell block sections of fat pad aspirates. Therefore, ultrastructural evaluation of fat pad aspirates by electron microscopy should be utilized, given its increased sensitivity for amyloid detection. This article demonstrates a simple and reproducible procedure for performing anterior fat pad aspiration for the detection of amyloid utilizing both Congo red staining of cell block sections and electron microscopy for ultrastructural identification.

Fat pad aspiration is a preferred, minimally invasive, and low cost approach as compared to other methods to detect amyloid for diagnosis of systemic amyloidosis. This video article demonstrates a procedural outline for performing fat pad aspiration with appropriate processing of the specimen for the optimal diagnostic outcome.

아밀로이드에 대한 앞쪽에 지방 패드의 FNA 공연 및 처리

Historically, heart, liver, and kidney biopsies were performed to demonstrate amyloid deposits in amyloidosis.  Since the clinical presentation of this ...

연구

의학

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells (ASCs)

The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix (ECM) remodeling. In vivo, ECM-rich environment consisting of fibrillar collagens provides a structural support to adipose tissues during the progression and regression of obesity. Physiological ECM remodeling mediated by matrix metalloproteinases (MMPs) plays a major role in regulating adipose tissue size and function1,2. The loss of physiological collagenolytic ECM remodeling may lead to excessive collagen accumulation (tissue fibrosis), macrophage infiltration, and ultimately, a loss of metabolic homeostasis including insulin resistance3,4. When a phenotypic change of the adipose tissue is observed in gene-targeted mouse models, isolating primary ASCs from fat depots for in vitro studies is an effective approach to define the role of the specific gene in regulating the function of ASCs. In the following, we define an immunomagnetic separation of Sca1high ASCs.

Immunomagnetic Separation of Fat Depot-specific Sca1high Adipose-derived Stem Cells ASCs

We present the techniques required to isolate the stromal vascular fraction (SVF) from mouse inguinal (subcutaneous) and perigonadal (visceral) adipose tissue depots to assess their gene expression and collagenolytic activity. This method includes the enrichment of Sca1high adipose-derived stem cells (ASCs) using immunomagnetic cell separation.

지방 창고 별 SCA1의 면역 자기 분리<sup&gt; 높은</sup&gt; 지방이 유래 줄기 세포 (의 ASC)

The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix ...

발생학

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly effective. This video describes the percutaneous biopsy technique using a modified Bergstr&#246;m needle to obtain skeletal muscle tissue samples from the vastus lateralis of human subjects. The Bergstr&#246;m needle consists of an outer cannula with a small opening (&#8216;window&#8217;) at the side of the tip and an inner trocar with a cutting blade at the distal end. Under local anesthesia and aseptic conditions, the needle is advanced into the skeletal muscle through an incision in the skin, subcutaneous tissue, and fascia. Next, suction is applied to the inner trocar, the outer trocar is pulled back, skeletal muscle tissue is drawn into the window of the outer cannula by the suction, and the inner trocar is rapidly closed, thus cutting or clipping the skeletal muscle tissue sample. The needle is rotated 90&#176;&#160;and another cut is made. This process may be repeated three more times. This multiple cutting technique typically produces a sample of 100-200 mg or more in healthy subjects and can be done immediately before, during, and after a bout of exercise or other intervention. Following post-biopsy dressing of the incision site, subjects typically resume their activities of daily living right away and can fully participate in vigorous physical activity within 48-72 hr. Subjects should avoid heavy resistance exercise for 48 hr to reduce the risk of herniation of the muscle through the incision in the fascia.

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

수정 된 버그 스트롬 기술을 이용하여 인간의 골격 근육 생검 절차

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

현지화, 식별 및 절제 쥣과 지방이 종점의

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and well tolerated technique. The Weil-Blakesley conchotome has a sharp biting tip with a 4 - 6 mm wide hollow. It is inserted through a 5 - 10 mm skin incision and can be maneuvered for controlled tissue penetration. The tip is opened and closed within the tissue and then rotated through 90 -180&#176; to cut the muscle. The amount of muscle obtained following repeated sampling can vary from 20 mg to 290 mg which can be processed for both histology and molecular studies. The wound needs to be kept dry and vigorous physical activity kept to a minimum for approximately 72 hr although normal levels of activity can restart immediately following the procedure. This procedure is safe and effective when close attention is paid to the selection of subjects, full asepsis and post procedure care. &#160;Both right and left vastus lateralis are suitable for biopsy dependent on participant preference.

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

This video demonstrates the technique of percutaneous muscle biopsy of the human vastus lateralis using the Weil-Blakesley conchotome.

사람의<em&gt; 광근의 Lateralis</em&gt; 웨일 - Blakesley Conchotome를 사용하여 골격 근육 생검

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and ...

Three-Dimensional Culture of Vascularized Thermogenic Adipose Tissue from Microvascular Fragments

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for the design of personalized microtissues for health screening and drug testing. Current strategies are often quite complex and fail to accurately fully depict the multicellular and functional properties of thermogenic adipose tissue. Microvascular fragments, small intact microvessels comprised of arteriole, venules, and capillaries isolated from adipose tissue, serve as a single autologous source of cells that enable vascularization and adipose tissue formation. This article describes methods for optimizing culture conditions to enable the generation of three-dimensional, vascularized, and functional thermogenic adipose tissues from microvascular fragments, including protocols for isolating microvascular fragments from adipose tissue and culture conditions. Additionally, best practices are discussed, as are techniques for characterizing the engineered tissues, and sample results from both rodent and human microvascular fragments are provided. This approach has the potential to be utilized for the understanding and development of treatments for obesity and metabolic disease.

Here, we present a detailed protocol outlining the use of microvascular fragments isolated from rodent or human fat tissue as a straightforward approach to engineer functional, vascularized beige adipose tissue.

Microvascular Fragments에서 Vascularized Thermogenic Adipose Tissue의 3차원 배양

Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for ...

생체공학

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.

지방 조직 유래 Stromal 혈관 분수 잠재적인 연골 재생을 위한 생산의 임상 프로토콜

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

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 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.

미니 지방 흡입 기술을 사용한 인간 피하 지방 조직 샘플링

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

Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images&#160;of an entire adipose tissue.

Due to the high lipid content, adipose tissue has been challenging to visualize using traditional histological methods. Adipo-Clear is a tissue clearing technique that allows robust labeling and high-resolution volumetric fluorescent imaging of adipose tissue. Here, we describe the methods for sample preparation, pretreatment, staining, clearing, and mounting for imaging.

Adipo-지우기: 조직 지방 조직의 3 차원 이미징 방법 삭제

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte ...

생물학

Human Adipose Tissue Micro-fragmentation for Cell Phenotyping and Secretome Characterization

In the past decade, adipose tissue transplants have been widely used in plastic surgery and orthopaedics to enhance tissue repletion and/or regeneration. Accordingly, techniques for harvesting and processing human adipose tissue have evolved in order to quickly and efficiently obtain large amounts of tissue. Among these, the closed system technology represents an innovative and easy-to-use system to harvest, process, and re-inject refined fat tissue in a short time and in the same intervention (intra-operatively). Adipose tissue is collected by liposuction, washed, emulsified, rinsed and minced mechanically into cell clusters of 0.3 to 0.8 mm. Autologous transplantation of mechanically fragmented adipose tissue has shown remarkable efficacy in different therapeutic indications such as aesthetic medicine and surgery, orthopedic and general surgery. Characterization of micro-fragmented adipose tissue revealed the presence of intact small vessels within the adipocyte clusters; hence, the perivascular niche is left unperturbed. These clusters are enriched in perivascular cells (i.e., mesenchymal stem cell (MSC) ancestors) and in vitro analysis showed an increased release of growth factors and cytokines involved in tissue repair and regeneration, compared to enzymatically derived MSCs. This suggests that the superior therapeutic potential of microfragmented adipose tissue is explained by a higher frequency of presumptive MSCs and enhanced secretory activity. Whether these added pericytes directly contribute to higher growth factor and cytokine production is not known. This clinically approved procedure allows the transplantation of presumptive MSCs without the need for expansion and/or enzymatic treatment, thus bypassing the requirements of GMP guidelines, and reducing the costs for cell-based therapies.

Here, we present human adipose tissue enzyme-free micro-fragmentation using a closed system device. This new method allows the obtainment of sub-millimeter clusters of adipose tissue suitable for in vivo transplantation, in vitro culture, and further cell isolation and characterization.

세포 표현형 및 분비 특성화를 위한 인간 지방 조직 미세 단편화

In the past decade, adipose tissue transplants have been widely used in plastic surgery and orthopaedics to enhance tissue repletion and/or regeneration. ...