サンプルおよび細胞培養プロシージャの確立の準備の彼の援助のマーティン ・ ダナーを認めた著者は、ドイツビスターフェルドベルンハルト Spezialchemie GmbH (ハンブルク、ドイツ)、継続的なサポートおよび議論のため特にロバート ・ Radsziwill に感謝したいと思います。これらの結果につながる研究は、欧州連合の第 7 フレームワーク プログラム (FP/2007-2013) ERC 助成契約名 340929 の下で欧州研究評議会から資金を受けています。

<ol>
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著者が明らかに何もありません。

複合構造物の設計には、ヤング率やサンプルの厚さなどの材料特性の簡単な調整ができます。PDMS のヤングは、2 つのコンポーネント間の混合比を変更することまたは別のシリコーンのエラストマー30,31を使用してブレンドの製造によって広い範囲で効果的に変更できます。説明の方法に限定されない PDMS の現在の調査で使用されるが、特に粘着性能は使用される特定のタイプによって強く決まります。このプロトコルの中で重要なステップは、複合膜 (図 1) の製造プロセスです。フィルムの厚さが大きく (図 5および図 6) 皮膚を含む、異なる基板上に薄膜の付着挙動で影響が示されました。膜厚だけでなく時間と硬化過程の温度32材料特性に影響します。したがって、ポリマー層の厚さとしてパラメーターは慎重に適応し、検証する必要があります。
薄膜の接着特性の解析を行った Ra 最大表面粗さの異なる 2 つのガラス基板を用いた垂直力付着測定 = 0.338 μ m (図 3)。一般に、粗さ影響大幅表面、特に弾性材料33,34の接着です。ガラスの粗さは異なるアスペリティ サイズ、高い粗さ値21を出展基板の作製を許可のサンドペーパーで研磨が簡単に変えることができます。たとえばまた、他の材料は基板15,35の生産のためエポキシ樹脂を使用できます。提案するプロトコルの変更の重要な戦略があります。たとえば、異なる表面自由エネルギーを示す基板が必要なのか地形は必要です。ここでは、引き剥がし応力と PDMS、SSA の製造された薄膜の分離の動作をカスタム セットアップ (巨視的密着性測定装置 (MAD、図 4)) を行った。36基板と圧子の光学配置は、測定結果の分析のための重要なステップです。したがって、チルト角度の調整は、ゴニオメータ、可能な限り正確に実行する必要があります。これは、水平方向の接触を達成するまで手動でフィルム表面と接触する基板を持って来ることによって十分な精度で実現できます。
現在のプロトコルのホールド時間が一定 1 秒後(図 5 と図 7)。特に、大まかな基板表面に弾性膜の接着性能の調査のためホールド時間の延長は、追加情報を示します。たとえば、保持時間の増加とともに引き剥がし応力の増加は、報告された8をされています。現在のプロトコルは、他の方法で実行される測定に加えてたとえば皮テスト実行でした、接着性能37のより包括的な調査を許可します。
複合膜柔らかい肌接着剤の厚さを調べた別のフィルムを展示しています (図 7) との接着。我々 の結果は公開されたデータ、監禁、すなわち、基板直径、膜厚の増加38,39 間の比率としてフィルム厚鉛引き剥がし応力の増加の減少を示すに沿っています。.これらの結果、図 7に示されているデータに基づいて、良好な密着性 p を約 100 μ m (約 60 μ m の厚約 40 μ m で PDMS 膜に適用の SSA 層の厚さ) の合計の厚さ複合膜に展示を終えてください。粗面のプロパティ。
次に、原始的な複合膜の生物学的特性に関連する実験を行ったし、プラズマ処理による複合膜 (図 8)。シリコン ・ エラストマーのプラズマ処理は表面および携帯電話の添付ファイルと携帯40,41を拡散促進の親水性の特性を高めるため多くの場合、汎用性の高い手法です。シリコーンは低毒性の高い biostability よく知られているが、残留モノマーまたは触媒細胞毒性42,43にもつながる生理的プロセスに影響を及ぼす可能性がありますがあります。で、我々 を観察した実験を行った未満 5% 細胞毒性指標とトリパン ブルー排除アッセイ LDH リリースを使用して。提案するプロトコル、全体の携帯電話人口の増殖解析 (図 9 b) 表面分析されているフォームをデタッチ細胞集合体を含みます。プロトコルの変更より差別化された結果を作り出すことができます。各サンプルでは、戸建の細胞凝集体を含む上清を別反応管を転送し、高分子表面から酵素によって削除されたセルと組み合わされていないでした。これは、表面に付着した細胞の正確な評価を許可し、最終的に細胞接着過程に及ぼすポリマーのより詳細な決定を明らかにします。に加えて免疫細胞化学的方法をここで提示、セルのイムノブロット法、蛋白質の表現の詳細な定量的評価を可能と調査のために収穫があります。
要約すると、高度な細胞培養研究用エラストマー複合薄膜の生産のための製造条件を設けています。また、これらの薄膜は、皮膚の粗さ、洗練されたデザインの皮膚接着剤を有効にするのに高い適応性を持っています。

このプロトコルでは、エラストマのフィルムの生産のための詳細な手順が掲載されています。広く利用可能なドクター刃技法は、複合薄膜の生産のために使用されています。製造技術は、大規模なこれらのフィルムの後続の生産を有効にする polyethylenterephtalate (ペット) 箔に行われています。このプロトコルの重点は、再現性、正確な複合膜の異なる層と最終的な複合材パッチの生物と接着特性の定量の製作の評価です。シリコーンのエラストマー ポリジメチルシロキサン (PDMS) は皮膚接着剤、マイクロ流体アプリケーション、その他の研究フィールド1,2,3 の生産を含む医療技術で広く使用されて ,4。最近、いわゆる柔らかい皮膚接着剤 (SSAs) PDMS の別のサブクラスは導入された、穏やかな皮膚の接着・非接着の特定されています。
シリコーン SSAs は、類似高分子シリカ5を補強の有無によって異なる官能基化ビニール エラストマーです。他の PDMS、SSA のようなヤング合わせることができる広い範囲で変調架橋剤濃度または硬化時間6,7,8。シリコーンエラストマーのヤング率の変化材料の接着特性に大きく影響し、表面9,10上で培養した原核生物と真核生物の細胞にも深遠な影響を持って,11. 細胞の生物学的レベルのそれを示した、真核細胞がマトリックスの弾力性の変調や表面9,10,12 の厚さに信号伝達レベルに対応 ,,1314。したがって、可変機械的性質を持つ高分子の細胞文化アプリケーションで広範な関心が存在します。重要なは、シリコーン系エラストマーの本質的に低表面エネルギーは、真核細胞の細胞培養に最適な条件を提供しません。酸素プラズマ処理は PDMS 低表面エネルギー、一時的に引き剥がし強度の向上につながるを高めるため広く使用されている手法は並列添付ファイル、拡散を推進中の分子の表面吸着を減少し、真核細胞15,16,17,18の増殖。
材料特性に加え表面の地形は細胞接着と接着剤 2 つ材料19,20,21,22相互に大きく影響します。表面粗さは、2 つのサーフェス間接触の形成に関するいくつかの効果: 高接触面積の削減にアスペリティをき裂伝ぱに及ぼす影響は、接着強度23を変えることができるだけでなく周囲の弾性エネルギーが格納されています。 24。人間の皮膚に粘着の接着は新興分野、例えば、創傷被覆材、心電図電極の固定または他ウェアラブル電子機器25,26,27、 28。表面地形に関連しての自己接着剤の接着性能を測定するには、通常接着測定8、21の粗さの度合いとガラス基板を使用できます。ここでは、2 枚のガラス基板は、高分子膜の接着特性を調査する選択されています。混合比の異なる PDMS 10 に 1 の重量部品の混合比の PDMS バッキング層と最初、複合フィルムは特徴付けられました。2 番目のステップでは、両方の成分の等しい重量量とサポート PDMS 膜上の膜厚を変化に接着 SSA 層が準備されました。

<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

このプロトコルの皮膚接着剤の開発、高度な細胞培養用エラストマ複合膜を作製する方法を提案する.(PDMS、柔らかい皮膚接着剤 (SSA))、2 つの異なるポリ-(dimethyl siloxanes) は、生物学的効果と接着特性の深さの調査に使用されています。複合フィルムは、柔軟なバッキング層・接着剤の上塗りで構成されます。両方のレイヤーは、ドクター刃アプリケーション技法によって製造されています。現在の調査で層の厚さの関数またはトップ層のヤング率の変化として複合膜の接着挙動を調べた。PDMS のヤングは、架橋剤の混合比にベースを変化させることにより変更されています。さらに、SSA の膜の厚さは約 16 μ m から約 320 μ m. 走査型電子顕微鏡 (SEM) に多様されている、光学顕微鏡は、厚さ測定のために使用されています。エラストマー フィルムとの接着は、膜厚、ポリマーと表面特性のヤング率によって強く決まります。したがって、表面粗さを示すガラス基板上にこれらのフィルムの通常の接着が検討されています。引き剥がし応力との分離作業は、シリコーンエラストマーの混合比に依存。
さらに、支持の支持層の上に置かれた柔らかい肌接着剤の厚さは、皮膚用パッチを生成するために多様されています。(混合比 10:1) PDMS 薄膜や SSA 映画 (混合比 50: 50) L929 マウス線維芽細胞の細胞接着、増殖細胞毒性が行われています。我々 は示されているここでは、初めて、両ポリマーの製造複合薄膜の横に並べて比較とその生物と粘着特性の調査を提示します。

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

異なるヤング係数または厚さを有する高分子薄膜複合構造体の製造のためのプロトコルが表示されます。フィルムは、高度な細胞培養研究または皮膚接着剤として生産されています。

細胞培養および皮膚のアプリケーション フィルム複合シリコン エラストマーを薄い: 製造とキャラクタリゼーション

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

Universidad San Sebastian

Research

JoVE Journal

Developmental Biology

10.6K ビュー.  INM - Leibniz Institute for New Materials. 異なるヤング係数または厚さを有する高分子薄膜複合構造体の製造のためのプロトコルが表示されます。フィルムは、高度な細胞培養研究または皮膚接着剤として生産されています。

ビデオ: 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.

機械的にダイナミックな細胞培養のための微細加工プラットフォーム

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

生物工学

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.

生体力学的解析と組織工学のための細胞由来の組織のリングを製作する監督セルラーセルフアセンブリ

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.

用絹フィルム培養システム in vitroで分析とマテリアルデザイン

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.

細胞間相互作用を操作するためのシリコンマイクロチップ

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

生物学

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.

微細加工チャネルにおける細胞移動の研究

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膜上のマイクロパターン細胞を伸ばす

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.

定量的、皮膚が創傷管理のためのコンフォーマルスキンのような電子システムの作製と評価

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空間の細胞培養のための細胞骨格などの生体適合性液晶エラストマー発泡体の合成

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.

デュアルのバイオミメティック基底膜として極薄 Porated 弾性ゲルの細胞培養

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.

マイクロマシンマイクロ流体プラットフォームにおける簡単な3次元スキンオンチップモデルの生成

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

細胞培養および皮膚のアプリケーションフィルム複合シリコンエラストマーを薄い: 製造とキャラクタリゼーション

要約

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この動画の章

機械的にダイナミックな細胞培養のための微細加工プラットフォーム

生体力学的解析と組織工学のための細胞由来の組織のリングを製作する監督セルラーセルフアセンブリ

用絹フィルム培養システム in vitroで分析とマテリアルデザイン

細胞間相互作用を操作するためのシリコンマイクロチップ

微細加工チャネルにおける細胞移動の研究

PDMS膜上のマイクロパターン細胞を伸ばす

定量的、皮膚が創傷管理のためのコンフォーマルスキンのような電子システムの作製と評価

3D空間の細胞培養のための細胞骨格などの生体適合性液晶エラストマー発泡体の合成

デュアルのバイオミメティック基底膜として極薄 Porated 弾性ゲルの細胞培養

マイクロマシンマイクロ流体プラットフォームにおける簡単な3次元スキンオンチップモデルの生成

細胞培養および皮膚のアプリケーション フィルム複合シリコン エラストマーを薄い: 製造とキャラクタリゼーション

要約

さらに動画を探す

この動画の章

機械的にダイナミックな細胞培養のための微細加工プラットフォーム

生体力学的解析と組織工学のための細胞由来の組織のリングを製作する監督セルラーセルフアセンブリ

用絹フィルム培養システム in vitroで分析とマテリアルデザイン

細胞間相互作用を操作するためのシリコンマイクロチップ

微細加工チャネルにおける細胞移動の研究

PDMS膜上のマイクロパターン細胞を伸ばす

定量的、皮膚が創傷管理のためのコンフォーマルスキンのような電子システムの作製と評価

3D空間の細胞培養のための細胞骨格などの生体適合性液晶エラストマー発泡体の合成

デュアルのバイオミメティック基底膜として極薄 Porated 弾性ゲルの細胞培養

マイクロマシンマイクロ流体プラットフォームにおける簡単な3次元スキンオンチップモデルの生成

細胞培養および皮膚のアプリケーションフィルム複合シリコンエラストマーを薄い: 製造とキャラクタリゼーション