Name: exploring the potential of mesenchymal stem cell sheet on the development of hepatocellular carcinoma in vivo
Uploaded: 2018-09-11T12:30:00
Description: Watch this Scientific Journal Video about Exploring the Potential of Mesenchymal Stem Cell Sheet on The Development of Hepatocellular Carcinoma In Vivo at JoVE.com

The authors would like to thank the staff of the Experimental Surgery and Animal Laboratory at the College of Medicine, King Saud University, for their cooperation and support-especially Hussain Almukhayzim and Hisham Aloudah. The authors would also like to acknowledge the media team at King Saud bin Abdul-Aziz University for preparing the visual material-especially Muath bin Ghannam and Abdulwahab Alsulami.

The protocol follows the animal care guidelines of King Saud University&#39;s ethical committee. Surgical procedures, anesthetics, and other medications used on animals are approved by King Saud University&#39;s ethical committee. All experimental work is performed by appropriately trained personnel.
1. Cell Sheet Construction
<ol>
	<li>Coat the culture dishes (3.5-cm temperature-responsive culture dishes) with undiluted fetal bovine serum (FBS) and ensure to cover all the surface of the dis...

<ol>
	<li>Marra, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+targets+and+oxidative+stress+biomarkers+in+hepatocellular+carcinoma:+an+overview.">Molecular targets and oxidative stress biomarkers in hepatocellular carcinoma: an overview.</a> Journal of Translational Medicine. 9, 171 (2011).</li><li>Yamada, K. M., Cukierman, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+tissue+morphogenesis+and+cancer+in+3D.">Modeling tissue morphogenesis and cancer in 3D.</a> Cell. 130 (4), 601-610 (2007).</li><li>Kushida, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decrease+in+culture+temperature+releases+monolayer+endothelial+cell+sheets+together+with+deposited+fibronectin+matrix+from+temperature-responsive+culture+surfaces.">Decrease in culture temperature releases monolayer endothelial cell sheets together with deposited fibronectin matrix from temperature-responsive culture surfaces.</a> Journal of Biomedical Materials Research. 45 (4), 355-362 (1999).</li><li>Okano, T., Yamada, N., Sakai, H., Sakurai, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+recovery+system+for+cultured+cells+using+plasma-treated+polystyrene+dishes+grafted+with+poly+(N-isopropylacrylamide).">A novel recovery system for cultured cells using plasma-treated polystyrene dishes grafted with poly (N-isopropylacrylamide).</a> Journal of Biomedical Materials Research Part A. 27 (10), 1243-1251 (1993).</li><li>Chamberlain, G., Fox, J., Ashton, B., Middleton, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concise+review:+mesenchymal+stem+cells:+their+phenotype,+differentiation+capacity,+immunological+features,+and+potential+for+homing.">Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing.</a> Stem Cells. 25 (11), 2739-2749 (2007).</li><li>Waterman, R. S., Tomchuck, S. L., Henkle, S. L., Betancourt, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+mesenchymal+stem+cell+(MSC)+paradigm:+polarization+into+a+pro-inflammatory+MSC1+or+an+Immunosuppressive+MSC2+phenotype.">A new mesenchymal stem cell (MSC) paradigm: polarization into a pro-inflammatory MSC1 or an Immunosuppressive MSC2 phenotype.</a> PLoS One. 5 (4), e10088 (2010).</li><li>Waterman, R. S., Henkle, S. L., Betancourt, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cell+1+(MSC1)-based+therapy+attenuates+tumor+growth+whereas+MSC2-treatment+promotes+tumor+growth+and+metastasis.">Mesenchymal stem cell 1 (MSC1)-based therapy attenuates tumor growth whereas MSC2-treatment promotes tumor growth and metastasis.</a> PLoS One. 7 (9), e45590 (2012).</li><li>Curtin, L. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+buprenorphine+in+a+postoperative+pain+model+in+rats.">Evaluation of buprenorphine in a postoperative pain model in rats.</a> Comparative Medicine. 59 (1), 60-71 (2009).</li><li>. Guidelines on Anesthesia and Analgesia in Rats - ULAM Guidelines and SOPs - Michigan Medicine Confluence Available from: <a target="_blank" href="https://wiki.med.umich.edu/display/ULAMGSOP/Guidelines+on+Anesthesia+and+Analgesia+in+Rats">https://wiki.med.umich.edu/display/ULAMGSOP/Guidelines+on+Anesthesia+and+Analgesia+in+Rats</a> (2017)</li><li>Alshareeda, A. T., Sakaguchi, K., Abumaree, M., Mohd Zin, N. K., Shimizu, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+of+cell+sheet+technique+on+the+development+of+hepatocellular+carcinoma+in+rat+models.">The potential of cell sheet technique on the development of hepatocellular carcinoma in rat models.</a> PLoS One. 12 (8), e0184004 (2017).</li><li>Russo, J., Russo, I. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atlas+and+histologic+classification+of+tumors+of+the+rat+mammary+gland.">Atlas and histologic classification of tumors of the rat mammary gland.</a> Journal of Mammary Gland Biology and Neoplasia. 5 (2), 187-200 (2000).</li><li>Lin, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+and+limitations+of+genetically+modified+mouse+models+in+drug+discovery+and+development.">Applications and limitations of genetically modified mouse models in drug discovery and development.</a> Current Drug Metabolism. 9 (5), 419-438 (2008).</li><li>Driscoll, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+preclinical+new+drug+research+program+of+the+National+Cancer+Institute.">The preclinical new drug research program of the National Cancer Institute.</a> Cancer Treatment Reports. 68 (1), 63-76 (1984).</li><li>Suzuki, R., Aruga, A., Kobayashi, H., Yamato, M., Yamamoto, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+novel+in+vivo+cancer+model+using+cell+sheet+engineering.">Development of a novel in vivo cancer model using cell sheet engineering.</a> Anticancer Research. 34 (9), 4747-4754 (2014).</li><li>Chen, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+the+cell+sheet+technique+in+tissue+engineering.">Application of the cell sheet technique in tissue engineering.</a> Biomedical Reports. 3 (6), 749-757 (2015).</li><li>Matsuura, K., Haraguchi, Y., Shimizu, T., Okano, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+sheet+transplantation+for+heart+tissue+repair.">Cell sheet transplantation for heart tissue repair.</a> Journal of Controlled Release. 169 (3), 336-340 (2013).</li><li>Matsuura, K., Shimizu, T., Okano, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+the+development+of+bioengineered+human+three-dimensional+vascularized+cardiac+tissue+using+cell+sheet+technology.">Toward the development of bioengineered human three-dimensional vascularized cardiac tissue using cell sheet technology.</a> International Heart Journal. 55 (1), 1-7 (2014).</li><li>Folkman, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+is+the+evidence+that+tumors+are+angiogenesis+dependent?.">What is the evidence that tumors are angiogenesis dependent?.</a> Journal of the National Cancer Institute. 82 (1), 4-6 (1990).</li></ol>

An extensive amount of research is dedicated to developing an adequate in vivo preclinical animal model that resembles human cancers. Currently, the major approaches used to create cancer animal models involve genetic engineering and cell transplantation11. Genetically modified animal models are good tools for the identification and validation of target genes, as well as understanding the molecular mechanisms underlying drug-induced toxicity. However, this model is associated with some li...

HCC is a primary cancer of the liver that is significantly associated with a poor prognosis. Annually, almost a half million new patients are diagnosed with HCC, representing 85% of liver cancer patients worldwide1.&#160;Hepatocarcinogenesis is not a single-form disease; rather,it is a collection of diseases that have different histopathological features and genetic and genomic variability, in addition to varied prognostic outcomes1. Therefore, the main challenges in the development of an effective therapeutic strategy for HCC are the limited knowledge of HCC biology and&#160;the lack of a suitable experimental animal model that can help understand this complex disease. An in vivo animal model that mimics human cancer is necessary for selecting candidate genes and identifying prognostic/predictive markers implicated in cancer induction, as well as for investigating different factors that may affect cancer responses to therapeutic agents.
In vitro cancer studies are still associated with major limitations. This is due to the fact that cancer cells lose many of their in vivo features when maintained in culture. The changes that occur to cells in vitro result from the absence of the whole tissue physiology in an ex vivo setting. Cancer cell-to-cell interaction (stromal, immune, vasculature, epithelial, etc.) within the tumor microenvironment reflects greatly on cancer cell characteristics2. The tumor microenvironment could alter cancer cell gene/protein expression and phenotypic characteristics, in addition to angiogenetic and metastatic potentials. The two-dimensional (2-D) in vitro culture system also lacks a suitable tissue matrix, which is necessary to regulate tumor progression. Thus, due to these limitations, in vivo models should always be utilized to support the preliminary findings of in vitro models. In this study, we use cell sheet technology to develop an in vivo animal model that elucidates the complete biological process underlying HCC.
More than a decade ago, Okano&#39;s laboratory established a novel method of tissue engineering based on cell sheet technology3. This technique utilizes a thermo-responsive culture plastic to allow reversible cell adhesion/detachment by controlling the surface hydrophobicity. This method allows a gentle harvesting of cultured cells in an intact three-dimensional (3-D) format (i.e.,cell sheet), with a well-kept extracellular matrix (ECM) and cell-to-cell interactions. The cell sheet technique requires the precoating of culture dishes with a temperature-responsive polymer poly(N-isopropylacrylamide) (PIPA Am), which is commercially available and ready for use. At a temperature below 20 &#176;C, PIPA Am polymers become hydrated and dissolve in aqueous solutions, whereas, at a higher temperature (37 &#176;C), the polymers become dehydrated and transform into a turbid precipitate. The polymer contains hydrophilic amide chains and hydrophobic side chains (isopropyl groups). At high temperatures, the Brownian movement of the water molecules is intensified, whereas, at a low temperature, the molecules of the water surrounding the isopropyl groups of the hydrated structure breakdown and the groups of hydrophobic isopropyl aggregate because of hydrophobic interactions. Therefore, the entire chain of the polymer aggregates and precipitates4.
In the presented study, this technique is employed to develop an HCC animal model using three different cell sheets. The first sheet employed consists of HCC cell line cells only, whereas the other two sheets consist of a combination of HCC cell line cells and MSCs from two different sources: bone marrow MSCs (BMMSCs) and umbilical cord MSCs (UCMSCs). MSCs are non-hematopoietic stromal cells that are capable of differentiating intocell derivatives of the mesenchymal lineage, including adipocytes, osteocytes, chondrocytes, and myocytes5. The reason we employ these cells when creating the cancer cell sheet is the inconsistent reporting on the effect of MSCs on cancers. It has been suggested that MSCs can have two distinct phenotypes: &#34;MSC1&#34;, a proinflammatory phenotype, and &#34;MSC2&#34;, an immunosuppressive phenotype6. MSCs express toll-like receptors (TLRs). TLR4 priming of MSCs increases their secretion of proinflammatory factors, whereas TLR3 priming increases their secretion of immunosuppressive factors6. An in vitro study of these two phenotypes reported that a co-culture of MSC1 with cancer cell lines attenuated cancer cell growth, while MSC2 co-culture had the opposite effect7. This implicates that MSCs can be either pro-cancer or anticancer, depending on their phenotype. Thus, in addition to developing the HCC animal model using cell sheet technology, we want to explore the effect of MSC transplants on tumor development, and whether using these cells will enhance or reduce the development of this model.

<table border="0" cellpadding="0" cellspacing="0" style="width:787px;" width="787">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:501px;">Reagents&#160;</td>
			<td></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">FBS&#160;</td>
			<td style="width:192px;">Gibco/Invitrogen</td>
			<td style="width:119px;">10270106</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">DMEM high glucose&#160;</td>
			<td style="width:192px;">Sigma</td>
			<td style="width:119px;">D5671-500ML</td>
			<td></td>
		</tr>
		<tr height="31">
			<td height="31" style="height:31px;width:309px;">Penicillin/streptomycin&#160;</td>
			<td style="width:192px;">Life Technology</td>
			<td style="width:119px;">15070063</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:309px;">Sterile physiologic saline</td>
			<td style="width:192px;">Sigma&#160;</td>
			<td style="width:119px;">S0817-1GA</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:309px;">Human HepG2 cell line</td>
			<td style="width:192px;">ATCC, USA</td>
			<td style="width:119px;">HB-8065</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:309px;">Human bone marrow MSCs cell line</td>
			<td style="width:192px;">PromoCell, USA</td>
			<td>C-12974</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:309px;">human umbilical cord tissue MSCs</td>
			<td style="width:192px;">PromoCell, USA&#160;</td>
			<td style="width:119px;">C-12971</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">Ketamine 50%</td>
			<td style="width:192px;">Rompun, Bayer</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">Xylazine 2%</td>
			<td style="width:192px;">Rompun</td>
			<td style="width:119px;">23076-35-9</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Alphadine&#174;&#160;solution.</td>
			<td style="width:192px;">Riyadh Pharma</td>
			<td style="width:119px;">LBL0816</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:501px;">Disposables:&#160;</td>
			<td></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:309px;">15mL Polypropylene High Clarity PP Centrifuge Tube</td>
			<td style="width:192px;">Falcon&#160;</td>
			<td style="width:119px;">352097</td>
			<td></td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:309px;">3.5 cm sterile UpCell culture dishes with the filter paper (membrane)</td>
			<td style="width:192px;">Sigma</td>
			<td style="width:119px;">174904-1CS</td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:309px;">100-1000 &#181;l &#160;Pipette Tips&#160;</td>
			<td style="width:192px;">Sigma</td>
			<td style="width:119px;">CLS4868-1000EA&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Basic Procedure Drape</td>
			<td style="width:192px;">Thermofisher</td>
			<td style="width:119px;">PMD5293.0</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:501px;">Equipment&#160;</td>
			<td></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">Plus pipette, variable volume</td>
			<td style="width:192px;">Eppendorf&#174;&#160;Research&#174;</td>
			<td style="width:119px;">Z683779-1EA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:309px;">Tissue culture incubator 37 &#176;C, 5% CO2</td>
			<td style="width:192px;">Any brand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:309px;">Biological safety cabinet</td>
			<td style="width:192px;">Any brand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:309px;">Tissue culture incubator 20 &#176;C, 5% CO2</td>
			<td style="width:192px;">Any brand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td colspan="2" height="22" style="height:22px;width:501px;">Sterile surgical tools and nude rats:&#160;</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td colspan="2" height="22" style="height:22px;width:501px;">Forceps</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td colspan="2" height="22" style="height:22px;width:501px;">Scissors</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td colspan="2" height="22" style="height:22px;width:501px;">scalpel&#160;</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">&#160;Nylon Suture&#160; 5-0</td>
			<td style="width:192px;">Accutome</td>
			<td style="width:119px;">AB-3854S</td>
			<td>Monofilament, Lancet</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">1 ml Tuberculin Syringes</td>
			<td style="width:192px;">Fisher Scientific</td>
			<td style="width:119px;">14-826-88</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:309px;">Nude rats&#160;</td>
			<td style="width:192px;">Charles river</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

exploring the potential of mesenchymal stem cell sheet on the development of hepatocellular carcinoma in vivo

An in vivo animal model that mimics human cancer could have various applications that deliver significant clinical information. The currently used techniques for the development of in vivo cancer models have considerable limitations. Therefore, in this study, we aim to implement cell sheet technology to develop an in vivo cancer model. Hepatocellular carcinoma (HCC) is successfully developed in nude rats using cell sheets created from HCC cell line cells. The cancer cell sheets are generated through intracellular adhesion and the formation of a stratified structure, controlled by the extracellular matrix. This allows for the HCC sheet transplantation into the liver and the creation of a tumor-bearing animal model within a month. In addition, the role of mesenchymal stem cells (MSC) in the development of this cancer model is investigated. In addition to the HCC cell line sheet, another two cell sheets are created: a sheet of HCC cells and bone marrow MSCs (BMMSCs) and a sheet of HCC cells and umbilical cord MSCs (UCMSCs). Sheets that have a combination of both HCC cells and MSCs are also capable of producing a tumor-bearing animal. However, the addition of MSCs reduces the size of the formed tumor, and this adverse effect on tumor development varies depending on the used MSCs' source. This indicates that a cell sheet made of certain MSC subtypes could be utilized in tumor management and control.

Tumorigenicity of Transplanted Cell Sheets in Rats:
One month after the transplantation, all transplanted cell sheets in rats&#39; livers have developed tumors (Figure 3). The average size of the developed tumors from HepG2, HepG2/BMMSC, and HepG2/UCMSC cell sheets were 4.5 cm, 4 cm, and 2.5 cm, respectively10.
<p class="jove_content" fo:keep-together.wit...

Watch this Scientific Journal Video about Exploring the Potential of Mesenchymal Stem Cell Sheet on The Development of Hepatocellular Carcinoma In Vivo at JoVE.com

Exploring the Potential of Mesenchymal Stem Cell Sheet on The Development of Hepatocellular Carcinoma In Vivo

Here, we present a protocol to develop an in vivo cancer model using cell sheet technology. Such a model could be very useful for the evaluation of anticancer therapeutics.

Exploring the Potential of Mesenchymal Stem Cell Sheet on The Development of Hepatocellular Carcinoma In Vivo

An in vivo animal model that mimics human cancer could have various applications that deliver significant clinical information. The currently used ...

Research

JoVE Journal

Bioengineering

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells hESCs and Induced Pluripotent Stem Cells iPSCs

We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to ...

Microinjection Wound Assay and In vivo Localization of Epidermal Wound Response Reporters in Drosophila Embryos.

Microinjection Wound Assay and In vivo Localization of Epidermal Wound Response Reporters in Drosophila Embryos.

The Drosophila embryo develops a robust epidermal layer that serves both to protect the internal cells from a harsh external environment as well as to ...

Procedure for the Development of Multi-depth Circular Cross-sectional Endothelialized Microchannels-on-a-chip

Efforts have been focused on developing in vitro assays for the study of microvessels because in vivo animal studies are more time-consuming, expensive, ...

Optimizing Attachment of Human Mesenchymal Stem Cells on Poly(&#949;-caprolactone) Electrospun Yarns

Optimizing Attachment of Human Mesenchymal Stem Cells on PolyÃƒÅ½Ã‚Âµ-caprolactone Electrospun Yarns

Research into biomaterials and tissue engineering often includes cell-based in vitro investigations, which require initial knowledge of the starting cell ...

Construction of Defined Human Engineered Cardiac Tissues to Study Mechanisms of Cardiac Cell Therapy

Human cardiac tissue engineering can fundamentally impact therapeutic discovery through the development of new species-specific screening systems that ...

Light-sheet Fluorescence Microscopy to Capture 4-Dimensional Images of the Effects of Modulating Shear Stress on the Developing Zebrafish Heart

The hemodynamic forces experienced by the heart influence cardiac development, especially trabeculation, which forms a network of branching outgrowths ...

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...

Patterning the Geometry of Human Embryonic Stem Cell Colonies on Compliant Substrates to Control Tissue-Level Mechanics

Human embryonic stem cells demonstrate a unique ability to respond to morphogens in vitro by self-organizing patterns of cell fate specification that ...

Isolation, Expansion, and Differentiation of Mesenchymal Stem Cells from the Infrapatellar Fat Pad of the Goat Stifle Joint

The IFP, present in the knee joint, serves as a promising source of MSCs. The IFP is an easily accessible tissue as it is routinely resected and discarded ...