Name: stabilizing hepatocellular phenotype using optimized synthetic surfaces
Uploaded: 2014-09-26T14:00:00
Description: Watch this Scientific Journal Video about Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces at JoVE.com

DCH, MB와 FK는 기금에 EPSRC 사후에 의해 지원되었다. BL-V와 DS는 각 MRC의 박사 studentships에 의해 지원되었다. KC는 영국 재생 의학 플랫폼에서 자금에 의해 지원되었다.

PHNGAD (폴리 [1,6-hexanodiol / 네오 펜틸 글리콜 / 디 (에틸렌 글리콜) -alt - 아 디프 산] 디올)의 1 합성 <img alt="반응식 1" fo:content-width="5in" src="/files/ftp_upload/51723/51723scheme1.jpg" width="500" /> 반응식 1 : PHNAGD의 합성 PHNAGD의 합성의 도식 표현.. PHNAGD는 1,6-Hexanodiol의 반응, 디 에틸렌 글리콜, n​​eoppentyl 글리콜 및 아 디프 산을 제조 하였다. PHNAGD, 폴리 [1,6-hexanodiol / 네오 펜틸 ?...

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줄기 세포로부터 간세포를 생성하는 데 사용되는 현재의 방법의 대부분은 동물 유래의 미정 행렬에 의존한다. 이러한 기판은 어플리케이션에 상당한 장벽을 나타내는 세포의 기능과 안정성에 영향을 미치는, 비싸고 매우 가변적 일 수있다. 따라서, 우리는 줄기 세포 유래 간세포의 문화를 지원하는 합성 물질에 대한 화면을 수행 하였다. 우리가 식별 한 견고한 간세포 분화 방식과 조합 PHNGAD, MDI ...

생물학적 물질은 널리 능성 줄기 세포의 한 유지 및 분화에 사용되어왔다. 가능하지만, 이러한 생물학적 기질은 종종 정의되지 않은 구성 요소의 무수가 포함되어 있습니다. 마트 리겔은 줄기 세포 배양 및 분화를위한 일반적으로 사용되는 기판이다. 불행하게도, 그 변수 조성물은 세포 표현형 및 기능에 영향을 미친다. 대안 이상의 정의 된 생물학적 매트릭스들의 다양한 2-7을 사용되었지만, 그들의 동물성 또는 나쁨 확장 그들에게 산업 제조에 부적합한 후보를 만든다. 따라서, 줄기 세포 연구의 주요 목표는 정의 된 구성과 신뢰할 수있는 성능으로 합성 대안의 식별이다. 정의되지 않은 세포 배양 기판의 한계를 극복하기위한 시도에서, 화학, 생물학 학제 간의 협력은 세포 표현형을 지원하는 능력과 합성 물질을 확인했다. 신디etic 기판은, 비용 효율적인 확장 성, 그리고 생체 내 환경을 흉내 낸, 복잡한 3 차원 구조로 제조 될 수있다. 이러한 특성으로 인해, 합성 기판은 널리 다양한 유형의 세포의 분화 8-10을 지원하고 구동하기 위해 사용되어왔다. 고급 및 높은 처리량 분석 대형 도서관에서 합성 물질의 신속한 심사를 촉진하고, 생물 의학 연구 및 개발 11-13에서 다양한 응용 프로그램과의 유연한 특성을 가진 새로운 물질을 전달했다. 높은 처리량, 고분자 마이크로 어레이 검사 기술을 활용, 우리는 빠른 속도로 인간의 줄기 세포 유래 간세포의 유지 보수에 적합한 간단한 폴리 우레탄 (PU134)를 확인했다. 이 중합체는 간세포 분화와 기능 14-16와 관련하여 동물 유래의 기재에 우수한 것으로 밝혀졌다. 우리는 이후 효과를 액세스하는 도포 조건, 지형 및 살균 처리를 최적화 한간세포 기능 및 수명을 안정화 중합체 성능. 이 세포 기반의 모델링 및 재생 의학 응용 프로그램에 대한 간세포 생물학의 기초를 이해 관련하여 중요한 의미를 갖는다. 여기에 설명 된 기술은 합성 중합체의 표면이 세포 표현형을 유지하도록 최적화 될 수있는 방법의 예를 나타낸다. 우리는 효율적인 무 혈청 간세포 분화 프로토콜이 기술의 조합이 체외 모델링 및 재생 의료에 사용하기위한 간세포의 생산 확장을 제공하는 잠재력을 가지고 있다고 믿는다.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Synthesis, preparation, coating and characterization of polymer PU134 coated coverslips</td>
		</tr>
		<tr>
			<td>Shaker</td>
			<td>Edmun B&#252;hler</td>
			<td>KS-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Irradiator</td>
			<td>CIS Biointernational</td>
			<td>IBL 637&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Spin coater</td>
			<td>Specialty Coating System</td>
			<td>&#160;P-6708</td>
			<td></td>
		</tr>
		<tr>
			<td>Scanning Electron Microscope&#160;</td>
			<td>Philips</td>
			<td>XL30CPSEM</td>
			<td></td>
		</tr>
		<tr>
			<td>Atomic Force Microscope</td>
			<td>DimensionV Nanoscope, VEECO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>p4-GLO CYP3A4</td>
			<td>Promega</td>
			<td>V8902</td>
			<td></td>
		</tr>
		<tr>
			<td>UV bulb</td>
			<td>ESCO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NanoScope analysis software</td>
			<td>VEECO</td>
			<td></td>
			<td>version 1.20</td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>Olympus</td>
			<td>TH45200</td>
			<td>Use Volocity 4 Software</td>
		</tr>
		<tr>
			<td>Tissue culture plates</td>
			<td>Corning, UK&#160;</td>
			<td>3527</td>
			<td></td>
		</tr>
		<tr>
			<td>glass slides</td>
			<td>Scientific Laboratory Supplies</td>
			<td>MIC3308</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethylene glycol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>93171</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;1,6-hexanediol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>240117</td>
			<td></td>
		</tr>
		<tr>
			<td>Neopentyl glycol</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>408255</td>
			<td></td>
		</tr>
		<tr>
			<td>Adipic acid</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>9582</td>
			<td></td>
		</tr>
		<tr>
			<td>anhydrous N,N-Dimethylformamide</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>227056</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethyl ether</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>676845</td>
			<td></td>
		</tr>
		<tr>
			<td>titanium (IV) butoxide&#160;</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>244112</td>
			<td></td>
		</tr>
		<tr>
			<td>1,4-butanediol&#160;</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>493732</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum oven</td>
			<td>Thermoscientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4,4&#8217;-Methylenebis(phenyl isocyanate)</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>101688</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofurane</td>
			<td>Sigma&#8211;Aldrich</td>
			<td>401757</td>
			<td></td>
		</tr>
		<tr>
			<td>Sputter coater</td>
			<td>Bal-Tec SCD 050</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Inmunostaining</td>
		</tr>
		<tr>
			<td>Phosphate buffer saline (-MgCl2, -CaCl2)</td>
			<td>Gibco</td>
			<td>10010031</td>
			<td>&#160;Store at room temperature</td>
		</tr>
		<tr>
			<td>PBST, PBS made up with 0.1% TWEEN 20&#160;</td>
			<td>&#160; Scientific Laboratory Supplies Ltd</td>
			<td>EC607&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>&#160; Scientific Laboratory Supplies Ltd</td>
			<td>CHE5010</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma-Aldrich, UK</td>
			<td>A7906</td>
			<td></td>
		</tr>
		<tr>
			<td>MOWIOL 488 DAPI</td>
			<td>Calbiochem</td>
			<td>475904</td>
			<td>Made up in Tris HCL and glycerol as per manufacturers instructions</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Cell culture and Functional assay</td>
		</tr>
		<tr>
			<td>CYP3A activity pGLO kit</td>
			<td>Promega</td>
			<td>V8902</td>
			<td></td>
		</tr>
		<tr>
			<td>Hepatozyme</td>
			<td>Gibco</td>
			<td>17705021</td>
			<td></td>
		</tr>
		<tr>
			<td>TryLE express</td>
			<td>Life Technologies</td>
			<td><a href="http://www.lifetechnologies.com/order/catalog/product/12604013">12604013</a></td>
			<td></td>
		</tr>
	</tbody>
</table>

stabilizing hepatocellular phenotype using optimized synthetic surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

고분자 용매 중합체 코팅 표면의 지형에 영향 폴리 우레탄 (134)은 단독으로 또는 톨루엔 또는 테트라 하이드로 퓨란 또는 디클로로 메탄과 유리 슬라이드가 다양한 제형으로 스핀 코팅과 조합하여, 클로로포름에 용해시켰다. 주사 전자 현미경 (SEM) 및 원자력 현미경 (AFM)을 (도 1) 중합체 코팅의 물리적 성질을 특성화하기 위해 사용되었다. 톨루엔, 클...

Watch this Scientific Journal Video about Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces at JoVE.com

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

이 문서는 장기적 폴리머 코팅 된 표면을 개발에 초점을 맞출 것이다, 줄기 세포의 안정적인 문화는 인간의 간세포를 산출했다.

최적화 된 합성 표면을 간세포 표현형 사용 안정화

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

Research

JoVE Journal

Chemistry

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

공유 결합은 자기 조립 단분자막의 접근 방식을 사용하여 표면에 BMP-2의 결합

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

연구

화학

Towards Biomimicking Wood: Fabricated Free-standing Films of Nanocellulose, Lignin, and a Synthetic Polycation

Nanocellulose, 리그닌의 날조 된 자립형 필름, 및 합성 폴리 양이온 : Biomimicking 나무를 향하여

Woody materials are comprised of plant cell walls that contain a layered secondary cell wall composed of structural polymers of polysaccharides and ...

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

In Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions

대량 선택 이온 소프트 랜딩에 의해 준비 잘 정의 된 표면의 제자리 SIMS 및 IR 분광법

Soft landing of mass-selected ions onto surfaces is a powerful approach for the highly-controlled preparation of materials that are inaccessible using ...

Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

고상 수지 기반의 방법론을 통해 비대칭 페로센 유래 바이오 복합체 시스템 용 합성 방법론

Early detection is a key to successful treatment of most diseases, and is particularly imperative for the diagnosis and treatment of many types of cancer. ...

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues

합성 단백질의 최적화 : Interpositional 종속 관계 확인이 구조적으로 표시 및 / 또는 기능적으로 연결된 잔류

Protein alignments are commonly used to evaluate the similarity of protein residues, and the derived consensus sequence used for identifying functional ...

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

를 사용하여 금 나노 입자의 효율적인 방사성 표지를위한 최적화 된 프로토콜<sup&gt; (125)</sup&gt; I-표시 아 지드 보철 그룹

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

Color Spot Test As a Presumptive Tool for the Rapid Detection of Synthetic Cathinones

색상 합성 Cathinones의 급속 한 탐지에 대 한 추정 도구로 자리 테스트

Synthetic cathinones are a large class of new psychoactive substances (NPS) that are increasingly prevalent in drug seizures made by law enforcement and ...

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) Mass Spectrometry

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight MALDI-TOF Mass Spectrometry

매트릭스를 통해 합성 고분자의 특성화 지원 레이저 탈 착 이온화 시간의 비행 (TOF MALDI) 질량 분석

There are many techniques that can be employed in the characterization of synthetic homopolymers, but few provide as useful of information for end group ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

빛 주도 분자 모터 표면에 단일 분자 이미징

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

A Facile Synthetic Method to Obtain Bismuth Oxyiodide Microspheres Highly Functional for the Photocatalytic Processes of Water Depuration

물 Depuration의 촉매 프로세스에 대 한 매우 기능적 비스무트 Oxyiodide 스피어를 손쉬운 합성 방법

Bismuth oxyhalide (BiOI) is a promising material for sunlight-driven-environmental photocatalysis. Given that the physical structure of this kind of ...