마틴 Danner 샘플 및 세포 문화 절차의 설립 준비에 그의 원조에 대 한 인정 이다. 저자는 베른하르트 Spezialchemie GmbH (함부르크, 독일), 지속적인 지원 및 토론에 대 한 특히 로버트 Radsziwill에 게 감사 하 고 싶습니다. 이러한 결과 연구는 유럽 연합의 7 차 프레임 워크 프로그램 (FP/2007-2013) ERC 부여 계약 n. 340929 아래 유럽 연구 위원회에서 자금을 받았다.

<ol>
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저자는 공개 없다.

복합 구조의 디자인에는 물질의 성질, 탄성 계수 또는 샘플의 두께 등의 간단한 조정 수 있습니다. PDMS의 영의 계수는 두 구성 요소 간의 혼합 비율을 변경 하거나 다른 실리콘 탄성 중합체30,31를 사용 하 여 혼합 제조 하 여 넓은 범위에서 효과적으로 바뀔 수 있다. 설명된 방법에 국한 되지 않습니다 PDMS의 현재 조사에 사용 하지만 특히 접착 성능 강하게 사용 특정 유형에 따라 다릅니다. 이 프로토콜 내에서 중요 한 단계는 복합 필름 (그림 1)의 제조 과정입니다. 그것은 그 두께 영화의 동작에 영향을 크게 접착 피부 (그림 5 및 그림 6)를 포함 하 여 다른 기판에 영화의 표시 했다. 필름 두께 뿐만 아니라 시간과 치료 과정 동안 온도 물성32을 영향을 줍니다. 따라서, 고분자 층의 두께 매개 변수 신중 하 게 적응 하 고 확인 해야 합니다.
박막의 접착 속성의 분석 직력 접착 측정 다른 표면 거칠기 Ra 최대 두 개의 유리 기판 사용 하 여 수행한 = 0.338 µ m (그림 3). 일반적으로, 거칠기에 미치는 영향 크게 표면, 특히 탄성 재료33,34의 접착. 유리의 거칠기 다른 asperity 크기, 따라서 수 있도록 높은 거칠기 값21전시 기판의 제조의 샌드 페이퍼로 연마 하 여 쉽게 변화 될 수 있다. 또한, 기타 자료, 예를 들어 에폭시 수 지 사용할 수 있습니다 기판15,35의 생산을 위해. 이 제시 프로토콜의 중요 한 수정 전략을 수 있습니다. 예를 들어 서로 다른 표면 자유 에너지를 전시 하는 기판은 필요한 또는 특정 topographies 있다면 필요. 여기, 풀에서 스트레스와 PDMS와 특수 제조 된 박막의 분리의 작품 사용자 설정 (거시적인 접착 력 측정 장치 (매드, 그림 4))으로 분석 되었다. 36 기판의 indenter 광학 정렬 측정 결과의 분석을 위한 중요 한 단계입니다. 따라서, 기울기 각도의 조정 가능한 정확한 각도 사용 하 여 수행 해야 합니다. 이 여 수동으로 필름 표면 접촉 기판 수평 접촉 달성 될 때까지 충분 한 정밀도를 얻을 수 있습니다.
현재 프로토콜에서 지속 시간 1 초 (그림 5 및 그림 7)에서 일정 유지 되었다. 거친 기판 표면에 탄력 있는 영화의 접착 성능 조사를 위해 특히 보류 시간 확장 추가 정보를 제공합니다. 예를 들어 보류 시간 증가 함께 풀 오프 스트레스 증가 보고8되었습니다. 현재 프로토콜을 다른 방법에서 수행 하는 측정 이외에 예 필 테스트 수 수행할 수, 접착 성능37의 포괄적인 조사를 허용.
복합 필름 다른 필름 소프트 스킨 접착제의 두께 결정 되었다 전시 (그림 7)의 접착 속성. 우리의 결과 게시 된 데이터, 그는 감 금, 즉, 기판 직경 및 필름 두께, 증가38,39 사이의 비율 풀에서 스트레스의 증가를 필름 두께의 감소를 보여주는 라인 . 이러한 결과 그림 7에 표시 된 데이터를 바탕으로, 우리는 결론을 약 100 µ m (약 40 μ m의 두께 가진 PDMS 영화에 적용 약 60 µ m의 특수 층의 두께)의 총 두께 가진 복합 영화 유리한 접착 p 전시 거친 표면에 roperties입니다.
다음, 깨끗 한 복합 영화에 생물 학적 특성에 관련 된 실험 수행 되었습니다 그리고 플라즈마 치료 복합 필름 (그림 8). 실리콘 탄성 체의 플라즈마 처리 표면 및 세포 부착 및 휴대40,41확산의 친수성 특성을 증가 하는 자주 적용, 다양 한 기법입니다. 실리콘 그들의 낮은 독성과 높은 biostability에 대 한 유명 하지만 잔여 단위체 또는 세포 독성42,43에 또한 지도 하는 생리 적 프로세스에 영향을 줄 수 있는 촉매를 포함 될 수 있습니다. 에 우리가 관찰 실시 실험 보다 5% 세포 독성 LDH 릴리스 표시기 및 Trypan 블루 제외 분석 결과 사용 하 여. 제시 프로토콜, 전체 세포 인구 세포 집계를 포함 한 표면 확산 분석 (그림 9B) 위해 분석 된 형태로 분리. 프로토콜의 보다 차별화 된 결과 생산할 수 있는. 각 샘플에 대 한 상쾌한 분리 된 세포 집계를 포함 하는 별도 반응 관에 전송 고 효소 폴리머 표면에서 제거 하는 세포와 결합 되지 않이 될 수 있습니다. 이 세포 표면에 부착 된의 정확한 평가 허용 하 고 결국 세포 접착 과정에는 폴리머의 영향의 더 상세한 결정을 공개 것입니다. 여기에 제시 된 immunocytochemical 방법, 이외에 셀 immunoblot 방법, 단백질 식의 구체적인된 양적 평가 수 있도록 조사에 대 한 수확 수 있습니다.
요약, 우리는 탄성 복합 박막 고급 셀 문화 연구에 응용 프로그램에 대 한 생산 제조 조건 확립. 또한, 이러한 박막 거칠음, 피부 접착제의 정교한 디자인을 사용 하면 피부에 높은 적응성을 소유한 다.

이 프로토콜에서 얇은 탄성 중합체 필름 생산을 위한 상세한 방법 제공 됩니다. 널리 의사 블레이드 기술은 복합 박막의 생산에 사용 되었습니다. 제조 방법 polyethylenterephtalate (애완 동물) 포 일, 대규모에서이 영화의 후속 생산 사용에 수행 되었습니다. 이 프로토콜의 중점은 재현성, 정확한 합성 영화의 다른 레이어와 최종 합성 패치의 생물 및 접착 속성의 결정의 제조의 평가 이다. 실리콘 엘라 스토 머 poly-(dimethylsiloxane) (PDMS)은 피부 접착제, 마이크로 응용 프로그램 및 추가 연구 분야1,2,3의 생산을 포함 하 여, 생물 의학 기술에 광범위 하 게 사용 ,4. 최근, PDMS, 부드러운 피부 접착제 (SSAs) 소위의 다른 서브 클래스 도입, 부드러운 피부 결합 및 결합 해제에 대 한 특정 있다.
실리콘 SSAs 실리 카5강화의 부재에 의해 유사한 중합체에서 서로 다른 기능성된 비닐 탄성 체입니다. 비슷한 다른 PDMS, SSA의 탄성 계수 수 적응 시킬 다양 한 변조 cross-linker 농도 또는 경화 시간6,,78. 이 실리콘 탄성 체의 탄성 계수에 크게 재료의 접착 특성에 영향을 미치는 변화와 간결한 및 진 핵 세포 표면9,10 에 교양에 깊은 결과가 또한 있다 , 11. 세포 생물학 수준에서 그것은 표시 했다, 진 핵 세포 신호 변환 수준에 매트릭스 탄성의 변조 또는 표면9,10,12의 두께에 대응 ,,1314. 따라서, 셀 문화 가변 기계적 특성을 가진 고분자의 애플 리 케이 션에 광범위 한 관심을 존재 한다. 중요 한 것은, 본질적으로 낮은 표면 에너지 기반 실리콘 탄성 체의 진 핵 세포의 세포 배양을 위한 최적의 조건을 제공 하지 않습니다. 산소 플라즈마 처리는 증가 PDMS 낮은 표면 에너지 일시적으로, 풀-오프 강도 향상을 선도 하는 널리 사용 되 기술 감소 첨부, 확산을 추진 하는 동시에 분자의 표면 흡착 하 고 진 핵 세포15,16,,1718의 확산.
재료 속성 뿐만 아니라 표면 지형을 크게 세포 접착 및 두 재료19,20,,2122사이 접착 상호 작용 영향을 줍니다. 표면 거칠기는 연락처 형성 두 표면 사이 여러 가지 효과: 접촉 지역, 높은의 감소 뿐만 아니라 균열 전파에 영향 접착 력23, 변경할 수 있는 asperities를 둘러싼 탄성 에너지 저장 24. 인간의 피부에 접착 필름의 접착은 한 신흥 응용 분야, 예를 들어, 상처 드레싱, ECG 전극의 고정 또는 다른 전자 장치를 착용 할 수 있는25,,2627, 28. 표면 지형에 관하여 자기-접착제의 접착 성능을 측정 하기 위해 거칠기의 학위를 변화와 함께 유리 기판 일반 접착 측정8,21에서 사용할 수 있습니다. 여기, 두 개의 유리 기판 고분자 필름의 접착 속성을 조사 선정 됐다. 10 1 중량 부품 PDMS 다른 혼합 비율 적용의 혼합 비율에 PDMS 백업 층으로 첫째, 복합 필름 특징 이었다. 두 번째 단계에서 접착제 특수 계층 동등한 무게 양의 두 구성 요소와 지원 PDMS 필름 위에 두께 변화를 준비 했다.

<table><tbody><tr><td>2-Propanol, 97%</td><td>Stockmeier Chemie</td><td>1000452610000</td><td>Isopropanol</td></tr><tr><td>Abrasive diamnod hand pad</td><td>Bohle</td><td>MO 5007522</td><td>Grit: 220</td></tr><tr><td>Accutase</td><td>Capricorn Scientific</td><td>ACC-1B</td><td></td></tr><tr><td>Albumin Fraktion V</td><td>Roth</td><td>0163.2</td><td>BSA</td></tr><tr><td>Alexa Fluor 488 Phalloidin</td><td>ThermoFischer Scientific</td><td>A12379</td><td>highly toxic</td></tr><tr><td>Aquamount</td><td>Polysciences</td><td>18606-20</td><td>water soluble mounting medium</td></tr><tr><td>CytoTox-ONE Homogeneous Membrane Integrity Assay</td><td>Promega</td><td>G7890</td><td></td></tr><tr><td>DPBS, without Ca&lt;sup&gt;2+&lt;/sup&gt;, Mg&lt;sup&gt;2+&lt;/sup&gt;</td><td>ThermoFischer Scientific</td><td>14190094</td><td></td></tr><tr><td>Fetal bovine serum gold</td><td>GE Health Care Life Science</td><td>A15-151</td><td>FBS</td></tr><tr><td>Goniometer OCA35</td><td>Dataphysics</td><td></td><td>for the determination of the static water contact angle</td></tr><tr><td>Hoechst Dye 33342</td><td>Sigma-Aldrich</td><td>B1155-100MG</td><td>bisBenzimide H 33342 trihydrochloride, highly toxic</td></tr><tr><td>Microscope Axiovert 25</td><td>Zeiss</td><td></td><td>Microscope used for cell culture documentation</td></tr><tr><td>Microscope Eclipse LV100ND</td><td>Nikon</td><td></td><td>Microscope used for film thickness determination</td></tr><tr><td>Paraformaldehyde, aqueous solution 16%</td><td>Electron Microscopy Sciences</td><td>RT 15710</td><td>electron microscopy grade</td></tr><tr><td>penicillin und streptomycin solution</td><td>Sigma-Aldrich</td><td>P4333-100ML</td><td></td></tr><tr><td>Phenom XL Scanning Electron Microscope (SEM)</td><td>Phenom</td><td></td><td></td></tr><tr><td>Poly-(vinyl alcohol) 4-88, MW 31000</td><td>Sigma-Aldrich</td><td>81381-1KG</td><td>Mowiol 4-88</td></tr><tr><td>Poly-dimethyl siloxanes, Sylgard 184</td><td>Dow Corning</td><td>(400)000108351397</td><td>PDMS</td></tr><tr><td>RPMI 1640 basal medium</td><td>ThermoFischer Scientific</td><td>21875034</td><td></td></tr><tr><td>soft skin adhesive (SSA)</td><td>Dow Corning</td><td>(400)000108251792</td><td>MG 7-9800 Soft Skin Adhesive (SSA)</td></tr><tr><td>speed mixer DAC 600.2 VAC-P</td><td>Hauschild</td><td></td><td></td></tr><tr><td>stylus profilomter</td><td>Zeiss</td><td></td><td>Model: SURFCOM 1500SD3</td></tr><tr><td>Tecan Infinite M200 pro</td><td>Tecan</td><td></td><td>fluorescence plate reader</td></tr><tr><td>Triton X 100</td><td>Calbiochem</td><td>648466</td><td></td></tr><tr><td>Trypan Blue solution</td><td>Sigma-Aldrich</td><td>T8154-100ML</td><td>highly toxic</td></tr><tr><td>Trypsin/EDTA solution</td><td>PAN-Biotech</td><td>P10-023500</td><td>0.05% Trypsin, 0.02% EDTA in PBS</td></tr><tr><td>UV glue</td><td>Bohle</td><td>BO MV76002</td><td>medium viscosity</td></tr></tbody></table>

thin film composite silicon elastomers for cell culture and skin applications: manufacturing and characterization

이 프로토콜에서 선물이 얇은 탄성 복합 필름 고급 셀 문화 응용 프로그램 및 피부 접착제의 개발에 대 한 조작 하는 방법. 두 개의 다른 폴 리-(dimethyl siloxanes) (PDMS와 소프트 스킨 접착제 (SSA)), 생물학적 효과 및 접착 특성의 깊이 조사에 대 한 사용 되었습니다. 유연한 백업 층 및 접착 최고 코팅 복합 필름에 의하여 이루어져 있다. 두 레이어 의사 블레이드 응용 프로그램 기술에 의해 제조 되어 있다. 현재 조사에 복합 필름의 접착 동작 레이어 두께의 함수 또는 상위 계층의 영의 모듈러스의 변형으로 조사 되었습니다. PDMS의 영의 계수는 crosslinker 혼합 비율에 기초를 변화 하 여 변경 되었습니다. 또한, 특수 필름의 두께 약 16 µ m에서 약 320 µ m. 스캐닝 전자 현미경 (SEM)으로 변화 되 고 광학 현미경 두께 측정을 위해 사용 되었습니다. 탄성 중합체 필름의 접착 속성 강하게 필름 두께, 중합체 및 표면 특성의 영의 계수에 따라 달라 집니다. 따라서, 부드럽고 거친 표면 전시 유리 기판에이 영화의 일반 접착 조사 되었습니다. 스트레스 풀 오프와 분리의 실리콘 탄성 체의 혼합 비율에 따라 달라 집니다.
또한, 지원 배경 레이어 위에 부드러운 피부 접착제의 두께 피부 응용 프로그램에 대 한 패치를 생성 하기 위하여 변화 되었다. 세포 독성, 확산 및 PDMS 영화 (혼합 비율 10:1)와 특수 필름 (비율 50: 50 혼합)에 L929 murine 섬유 아 세포의 세포 접착 실시 되었습니다. 우리는 여기에 표시 된, 처음으로, 두 고분자의 제조 복합 박막의 측면에서 비교 하 고 그들의 생물 학적 및 접착 속성의 조사를 제시.

Watch this Scientific Journal Video about Thin Film Composite Silicon Elastomers for Cell Culture and Skin Applications: Manufacturing and Characterization at JoVE.com

Thin Film Composite Silicon Elastomers for Cell Culture and Skin Applications: Manufacturing and Characterization

다른 영의 계수 또는 두께 가진 고분자 박막 복합 구조의 제조 프로세스에 대 한 프로토콜 제공 됩니다. 영화는 고급 셀 문화 연구에 대 한 또는 피부 접착제로 생산 됩니다.

세포 배양 및 피부 응용 프로그램에 대 한 영화 복합 실리콘 탄성 체를 얇은: 제조 및 특성

In this protocol, we present methods to fabricate thin elastomer composite films for advanced cell culture applications and for the development of skin ...

thin-film-composite-silicon-elastomers-for-cell-culture-skin

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JoVE Journal

Developmental Biology

10.6K 조회수.  INM - Leibniz Institute for New Materials. 다른 영의 계수 또는 두께 가진 고분자 박막 복합 구조의 제조 프로세스에 대 한 프로토콜 제공 됩니다. 영화는 고급 셀 문화 연구에 대 한 또는 피부 접착제로 생산 됩니다.

비디오: Thin Film Composite Silicon Elastomers for Cell Culture and Skin Applications: Manufacturing and Characterization

Microfabricated Platforms for Mechanically Dynamic Cell Culture

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or fundamental cell biology studies is limited by current bioreactor technologies, which cannot simultaneously apply a variety of mechanical stimuli to cultured cells. In order to address this issue, we have developed a series of microfabricated platforms designed to screen for the effects of mechanical stimuli in a high-throughput format. In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and further demonstrate how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

기계 다이나믹 셀 문화 Microfabricated 플랫폼

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or ...

연구

생체공학

Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do not have suitable autologous donor vessels due to disease progression or previous harvest. The aim of vascular tissue engineering is to develop a suitable alternative source for these bypass grafts. In addition, engineered vascular tissue may prove valuable as living vascular models to study cardiovascular diseases. Several promising approaches to engineering blood vessels have been explored, with many recent studies focusing on development and analysis of cell-based methods.2-5 Herein, we present a method to rapidly self-assemble cells into 3D tissue rings that can be used in vitro to model vascular tissues.
To do this, suspensions of smooth muscle cells are seeded into round-bottomed annular agarose wells. The non-adhesive properties of the agarose allow the cells to settle, aggregate and contract around a post at the center of the well to form a cohesive tissue ring.6,7 These rings can be cultured for several days prior to harvesting for mechanical, physiological, biochemical, or histological analysis. We have shown that these cell-derived tissue rings yield at 100-500 kPa ultimate tensile strength8 which exceeds the value reported for other tissue engineered vascular constructs cultured for similar durations (&lt;30 kPa).9,10 Our results demonstrate that robust cell-derived vascular tissue ring generation can be achieved within a short time period, and offers the opportunity for direct and quantitative assessment of the contributions of cells and cell-derived matrix (CDM) to vascular tissue structure and function.

This article outlines a versatile method to create cell-derived tissue rings by cellular self-assembly. Smooth muscle cells seeded into ring-shaped agarose wells aggregate and contract to form robust three-dimensional (3D) tissues within 7 days. Millimeter-scale tissue rings are conducive to mechanical testing and serve as building blocks for tissue assembly.

자기 조립 셀룰러이 Biomechanical 분석 및 조직 공학을위한 셀 유래 조직 반지를 조작하는 감독

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do ...

Silk Film Culture System for in vitro Analysis and Biomaterial Design

Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment 1,2 . These materials are desirable because they possess highly controllable dimensional and material characteristics, are biocompatible and promote cell adhesion, can be modified through topographic patterning or by chemically altering the surface, and can be used as a depot for biologically active molecules for drug delivery related applications 3-8 . In addition, silk films are relatively straightforward to custom design, can be designed to dissolve within minutes or degrade over years in vitro or in vivo, and are produce with the added benefit of being transparent in nature and therefore highly suitable for imaging applications 9-13. The culture system methodology presented here represents a scalable approach for rapid assessments of cell-silk film surface interactions. Of particular interest is the use of surface patterned silk films to study differences in cell proliferation and responses of cells for alignment 12,14 . The seeded cultures were cultured on both micro-patterned and flat silk film substrates, and then assessed through time-lapse phase-contrast imaging, scanning electron microscopy, and biochemical assessment of metabolic activity and nucleic acid content. In summary, the silk film in vitro culture system offers a customizable experimental setup suitable to the study of cell-surface interactions on a biomaterial substrate, which can then be optimized and then translated to in vivo models. Observations using the culture system presented here are currently being used to aid in applications ranging from basic cell interactions to medical device design, and thus are relevant to a broad range of biomedical fields.

Silk Film Culture System for in vitro Analysis and Biomaterial Design

Silk films are a novel class of biomaterials readily customizable for an array of biomedical applications. The presented silk film culture system is highly adaptable to a variety of in vitro analyses. This system represents a biomaterial design platform offering in vitro optimization before direct translation to in vivo models.

위한 실크 영화 문화 시스템 체외에서 분석 및 Biomaterial 디자인

Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment ...

Silicon Microchips for Manipulating Cell-cell Interaction

The role of the cellular microenvironment is recognized as crucial in determining cell fate and function in virtually all mammalian tissues from development to malignant transformation.&nbsp; In particular, interaction with neighboring stroma has been implicated in a plethora of biological phenomena; however, conventional techniques limit the ability to interrogate the spatial and dynamic elements of such interactions.
In Micromechanical Reconfigurable Culture (RC), we employ a micromachined silicon substrate with moving parts to dynamically control cell-cell interactions through mechanical repositioning. Previously, this method has been applied to investigate intercellular communication in co-cultures of hepatocytes and non-parenchymal cells, demonstrating time-dependent interactions and a limited range for soluble signaling 1.
Here, we describe in detail the preparation and use of the RC system. We begin by demonstrating the handling of the device parts using tweezers, including actuating between the gap and contact configurations (cell populations separated by a narrow 80-&micro;m gap, or in direct intimate contact). Next, we detail the process of preparing the substrates for culture, and the multi-step cell seeding process required for obtaining confluent cell monolayers. Using live microscopy, we then illustrate real-time manipulation of cells between the different possible experimental configurations. Finally, we demonstrate the steps required in order to regenerate the device surface for reuse: toluene and piranha cleaning, polystyrene coating, and oxygen plasma treatment.

This article describes an experimental approach for dynamic regulation of cell-cell interactions between adherent cells on a micrometer scale. Manipulation of intercellular communication between hepatocytes and stromal cell is demonstrated. The developed platform enables investigation of cell-cell interactions in a variety of biological processes, including development and pathogenesis.

세포 세포 상호 작용을 조작에 대한 실리콘 마이크로 칩

The role of the cellular microenvironment is recognized as crucial in determining cell fate and function in virtually all mammalian tissues from ...

생물학

Study of Cell Migration in Microfabricated Channels

The method described here allows the study of cell migration under confinement in one dimension. It is based on the use of microfabricated channels, which impose a polarized phenotype to cells by physical constraints. Once inside channels, cells have only two possibilities: move forward or backward. This simplified migration in which directionality is restricted facilitates the automatic tracking of cells and the extraction of quantitative parameters to describe cell movement. These parameters include cell velocity, changes in direction, and pauses during motion. Microchannels are also compatible with the use of fluorescent markers and are therefore suitable to study localization of intracellular organelles and structures during cell migration at high resolution. Finally, the surface of the channels can be functionalized with different substrates, allowing the control of the adhesive properties of the channels or the study of haptotaxis. In summary, the system here described is intended to analyze the migration of large cell numbers in conditions in which both the geometry and the biochemical nature of the environment are controlled, facilitating the normalization and reproducibility of independent experiments.

A quantitative method to study spontaneous migration of cells in a one-dimensional confined microenvironment is described. This method takes advantage of microfabricated channels and can be used to study migration of large number of cells under different conditions in single experiments.

미세 채널에서 세포의 이동에 관한 연구

The method described here allows the study of cell migration under confinement in one dimension. It is based on the use of microfabricated channels, which ...

Stretching Micropatterned Cells on a PDMS Membrane

Mechanical forces exerted on cells and/or tissues play a major role in numerous processes. We have developed a device to stretch cells plated on a PolyDiMethylSiloxane (PDMS) membrane, compatible with imaging. This technique is reproducible and versatile. The PDMS membrane can be micropatterned in order to confine cells or tissues to a specific geometry. The first step is to print micropatterns onto the PDMS membrane with a deep UV technique. The PDMS membrane is then mounted on a mechanical stretcher. A chamber is bound on top of the membrane with biocompatible grease to allow gliding during the stretch. The cells are seeded and allowed to spread for several hours on the micropatterns. The sample can be stretched and unstretched multiple times with the use of a micrometric screw. It takes less than a minute to apply the stretch to its full extent (around 30%). The technique presented here does not include a motorized device, which is necessary for applying repeated stretch cycles quickly and/or computer controlled stretching, but this can be implemented. Stretching of cells or tissue can be of interest for questions related to cell forces, cell response to mechanical stress or tissue morphogenesis. This video presentation will show how to avoid typical problems that might arise when doing this type of seemingly simple experiment.

This manuscript presents a technique to apply or release forces on adherent cells or tissues using unidirectional stretching.

PDMS 막에 마이크로 패턴 세포를 스트레칭

Mechanical forces exerted on cells and/or tissues  play a major role in numerous processes. We have developed a device to stretch  cells plated on a ...

Fabrication and Characterization of a Conformal Skin-like Electronic System for Quantitative, Cutaneous Wound Management

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of cutaneous wound healing on the skin. Existing methods, however, still rely on visual inspections through various microscopic tools and devices that normally include high-cost, sophisticated systems and require well trained personnel for operation and data analysis. Here, we describe methods and protocols to fabricate a conformal, skin-like electronics system that enables conformal lamination to the skin surface near the wound tissues, which provides recording of high fidelity electrical signals such as skin temperature and thermal conductivity. The methods of device fabrication provide details of step-by-step preparation of the microelectronic system that is completely enclosed with elastomeric silicone materials to offer electrical isolation. The experimental study presents multifunctional, biocompatible, waterproof, reusable, and flexible/stretchable characteristics of the device for clinical applications. Protocols of clinical testing provide an overview and sequential process of cleaning, testing setup, system operation, and data acquisition with the skin-like electronics, gently mounted on hypersensitive, cutaneous wound and contralateral tissues on patients.

This article presents methods to fabricate and characterize a conformal, skin-like electronic system and protocols for the use in clinical applications, particularly on cutaneous wound management.

양적, 피부가 상처 관리를위한 등각 피부와 같은 전자 시스템의 제조 및 특성

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of ...

Synthesis of Biocompatible Liquid Crystal Elastomer Foams as Cell Scaffolds for 3D Spatial Cell Cultures

Here, we present a step-by-step preparation of a 3D, biodegradable, foam-like cell scaffold. These scaffolds were prepared by cross-linking star block co-polymers featuring cholesterol units as side-chain pendant groups, resulting in smectic-A (SmA) liquid crystal elastomers (LCEs). Foam-like scaffolds, prepared using metal templates, feature interconnected microchannels, making them suitable as 3D cell culture scaffolds. The combined properties of the regular structure of the metal foam and of the elastomer result in a 3D cell scaffold that promotes not only higher cell proliferation compared to conventional porous templated films, but also better management of mass transport (i.e., nutrients, gases, waste, etc.). The nature of the metal template allows for the easy manipulation of foam shapes (i.e., rolls or films) and for the preparation of scaffolds of different pore sizes for different cell studies while preserving the interconnected porous nature of the template. The etching process does not affect the chemistry of the elastomers, preserving their biocompatible and biodegradable nature. We show that these smectic LCEs, when grown for extensive time periods, enable the study of clinically relevant and complex tissue constructs while promoting the growth and proliferation of cells.

This study presents a methodology to prepare 3D, biodegradable, foam-like cell scaffolds based on biocompatible side-chain liquid crystal elastomers (LCEs). Confocal microscopy experiments show that foam-like LCEs allow for cell attachment, proliferation, and the spontaneous alignment of C2C12s myoblasts.

3D 공간 세포 배양을위한 세포 발판으로 생체 적합성 액정 엘라스토머 거품의 합성

Here, we present a step-by-step preparation of a 3D, biodegradable, foam-like cell scaffold. These scaffolds were prepared by cross-linking star block ...

Ultrathin Porated Elastic Hydrogels As a Biomimetic Basement Membrane for Dual Cell Culture

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies of endothelial-epithelial bilayers have traditionally relied on permeable support models that enable bilayer culture, but permeable supports are limited in their ability to replicate the diversity of human basement membranes. In contrast, hydrogel models that require chemical synthesis are highly tunable and allow for modifications of both the material stiffness and the biochemical composition via incorporation of biomimetic peptides or proteins. However, traditional hydrogel models are limited in functionality because they lack pores for cell-cell contacts and functional in vitro migration studies. Additionally, due to the thickness of traditional hydrogels, incorporation of pores that span the entire thickness of hydrogels has been challenging. In the present study, we use poly-(ethylene-glycol) (PEG) hydrogels and a novel zinc oxide templating method to address the previous shortcomings of biomimetic hydrogels. As a result, we present an ultrathin, basement membrane-like hydrogel that permits the culture of confluent cellular bilayers on a customizable scaffold with variable pore architectures, mechanical properties, and biochemical composition.

Current bilayer culture models do not allow for functional in vitro studies that mimic in vivo microenvironments. Using polyethylene glycol and a zinc oxide templating method, this protocol describes the development of an ultrathin biomimetic basement membrane with tunable stiffness, porosity, and biochemical composition that closely mimics in vivo extracellular matrices.

듀얼에 대 한 생체 모방 지하실 멤브레인으로 ultrathin Porated 탄성 Hydrogels 세포 배양

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies ...

Generation of a Simplified Three-Dimensional Skin-on-a-chip Model in a Micromachined Microfluidic Platform

This work presents a new, cost-effective, and reliable microfluidic platform with the potential to generate complex multilayered tissues. As a proof of concept, a simplified and undifferentiated human skin containing a dermal (stromal) and an epidermal (epithelial) compartment has been modelled. To accomplish this, a versatile and robust, vinyl-based device divided into two chambers has been developed, overcoming some of the drawbacks present in microfluidic devices based on polydimethylsiloxane (PDMS) for biomedical applications, such as the use of expensive and specialized equipment or the absorption of small, hydrophobic molecules and proteins. Moreover, a new method based on parallel flow was developed, enabling the in situ deposition of both the dermal and epidermal compartments. The skin construct consists of a fibrin matrix containing human primary fibroblasts and a monolayer of immortalized keratinocytes seeded on top, which is subsequently maintained under dynamic culture conditions. This new microfluidic platform opens the possibility to model human skin diseases and extrapolate the method to generate other complex tissues.

Here, we present a protocol to generate a three-dimensional simplified and undifferentiated skin model using a micromachined microfluidic platform. A parallel flow approach allows the in situ deposition of a dermal compartment for the seeding of epithelial cells on top, all controlled by syringe pumps.

마이크로머신 마이크로유체 플랫폼의 간소화된 3차원 스킨 온-어칩 모델 생성

This work presents a new, cost-effective, and reliable microfluidic platform with the potential to generate complex multilayered tissues. As a proof of ...