1. The study tested the hypothesis that pre-seeding endothelial cells (ECs) and fibroblasts (FBs) prior to cardiomyocytes (CMs) would create a microenvironment to support cardiac tissue development.
2. Results showed that ECs seeded two days before CMs promoted more elongated CM morphology. Collagen and laminin production increased over time in EC-FB co-cultures.
3. DNA content and viability remained stable in EC-FB co-cultures, while total collagen levels significantly increased from day 3 to day 10.
Characterization of embryoid bodies formed with different protocols 使用不同培養方式形...Honey Cheng
That's part of my first year master researching in 2011, National Chung Hsing-University, Taiwan. Mice embryonic stem cells differentiating with embryoid bodies in a unattached formed, so I summarized a slides review. 這是2011年在中興就讀研究所第一年時所研究的方向. 胚胎幹細胞能夠在懸浮狀態形成類胚體與分化, 所以為此我整理了一份簡報, 介紹不同方式形成類胚體之特性.
Thanksgiving is an American Holiday where people set aside a day to remember what God has done for them over the past one year and praise Him and give Him thanks for the works He has done in their lives. This game plays on the idea of looking forward yet being able to recognize and picture what is behind. It serves as a metaphor for us, also looking forward to the future, but taking some time to picture and recognize what God has done in the past."
Characterization of embryoid bodies formed with different protocols 使用不同培養方式形...Honey Cheng
That's part of my first year master researching in 2011, National Chung Hsing-University, Taiwan. Mice embryonic stem cells differentiating with embryoid bodies in a unattached formed, so I summarized a slides review. 這是2011年在中興就讀研究所第一年時所研究的方向. 胚胎幹細胞能夠在懸浮狀態形成類胚體與分化, 所以為此我整理了一份簡報, 介紹不同方式形成類胚體之特性.
Thanksgiving is an American Holiday where people set aside a day to remember what God has done for them over the past one year and praise Him and give Him thanks for the works He has done in their lives. This game plays on the idea of looking forward yet being able to recognize and picture what is behind. It serves as a metaphor for us, also looking forward to the future, but taking some time to picture and recognize what God has done in the past."
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
Rotator cuff repair using a stem cell approachZakary Bondy
This presentation communicates current methods for rotator cuff repair mainly focusing on mesenchymal and tendon-derived stem cells. It looks to expand on future research in this field by communicating a future experiment to expand on current knowledge of tendon-derived stem cells.
Indian Dental Academy: will be one of the most relevant and exciting training center with best faculty and flexible training programs for dental professionals who wish to advance in their dental practice,Offers certified courses in Dental implants,Orthodontics,Endodontics,Cosmetic Dentistry, Prosthetic Dentistry, Periodontics and General Dentistry.
Neuromics' is a leader in providing Biopharmas, Academic and Government with CFR compliant 2 and 3-D human primary cell assays, media and supplements for discovery. We also provide antibodies, proteins/growth factors, apoptosis kits and genetic engineering/manipulation tools.
Stem Cell Technology and its Clinical ApplicationDr. Barkha Gupta
Dr. Barkha Gupta has been teaching Veterinary Biochemistry as well as clinical physiology at CVAS, Udaipur and PGIVER, Jaipur. She has earlier served in various capacities in the Department of Animal Husbandry, Govt. of Rajasthan. She has several publications and awards to her credit. She is the PI of M-RAJUVAS Android Educational Mobile Application for Veterinary and Animal Sciences and Kiosk Information System for Farmers/Livestock Owners. Dr. Gupta is also IFBA Certified Professional.
Stem cell therapy for the bladder has been conducted mainly on an experimental basis in the areas of bladder dysfunction. The therapeutic efficacy of stem cells was originally thought to be derived from their ability to differentiate into various cell types. For more details visit: http://www.cryobanksindia.com/moms-corner/case-studies/
Stem cell therapy for the bladder has been conducted mainly on an experimental basis in the areas of bladder dysfunction. The therapeutic efficacy of stem cells was originally thought to be derived from their ability to differentiate into various cell types. For more details visit: http://www.cryobanksindia.com/moms-corner/case-studies/
1. Cell organization in co-cultures of cardiac cells and analysis of ECM synthesis
Assaf Shemesh
Supervisor: Prof. Smadar Cohen, Yulia Sapir, Dr. Emil Ruvinov
The Faculty of Engineering Sciences
Avram and Stella Goldstein-Goren
Department of Biotechnology Engineering
Abstract
Native heart tissue is composed of several cell types – mostly cardiomyocytes (CMs), fibroblasts (FBs) and endothelial cells (ECs). In this
project I tested a hypothesis that pre-seeding of ECs/FBs, 2 days prior to CMs, would lead to a microenvironment appropriate for the
development of a cardiac tissue. My results show that collagen and laminin are produced in the culture and that CMs display a more
developed morphology when seeded onto an already established array of ECs. Future plans will include tri-cell (ECs, FBs and CMs)
cardiac culture experiments with several seeding sequences and more detailed ECM analysis (i.e. different collagen types).
Introduction
In native cardiac tissue, FBs secrete the extra cellular matrix (ECM)
that form the connective tissue, ECs comprise the blood vessels,
and CMs form the beating muscle (1). Complex interactions
between those cells regulate their function and survival. Cardiac
cells are supported in their native state by an intricate network of
connective fibers and capillaries (2). In the lab, the cells are seeded
into macroporous alginate scaffolds. The scaffolds incorporate cell
adhesion peptides – RGD and HBP (heparin binding peptide) (3).
The cells then begin to secrete ECM and basal lamina components
of their own.
CMECFB
Alginate scaffold with
adhesion peptides
Cells that form the cardiac tissue
My hypothesis was that seeding ECs, 2 days before CMs, would
promote the development of a microenvironment (ECM and
vessel network), enabling the installment of CMs and their
maturation into a cardiac tissue.
Project Objectives
The long term objective is to create vascularized cardiac tissue.
Short term goals are:
• Finding the optimal seeding strategy for various cell co-cultures
in RGD/HBP adhesive alginate scaffolds.
• Evaluating cardiac tissue formation as well as analysis of ECM
components.
Materials and Methods
Alginate scaffolds decorated with RGD and HBP peptides
Immunofluorescence
Confocal
microscopy
Cell metabolic activity/viability– Alamar blue
DNA quantification – Hoechst 33258
Total collagen – Sircol colorimetric assay
Results
EC – CM co-culture
A B
Figure 1: Laser scanning confocal imaging of EC-CM co-culture, 3
days post seeding. Immunostaining for α - actinin (green, CMs),
CD-31 (red, ECs) and DAPI (blue, nuclei). (A) ECs seeded two
days before CMs. (B) ECs and CMs seeded simultaneously. (C)
ECs seeded alone. Bar=100 µm.
References
1. Iyer R.K., Chiu L.L.Y., and Radisic M. (2008), Microfabricated poly (ethylene
glycol) templates enable rapid screening of triculture conditions for cardiac tissue
engineering, J of Biomedical Materials Research Part A, Vol. 89,pp. 616-631.
2. Narmoneva D.A., Vukmirovic R., Davis M.E., Kamm R.D., and Lee R.T. (2004),
Endothelial cells promote cardiac myocyte survival and spatial reorganization,
Circulation, Vol. 110, no. 8, pp. 962-968.
3. Sapir Y., Kryukov O., and Cohen S. (2010), Integration of multiple cell-matrix
interactions into alginate scaffolds for promoting cardiac tissue regeneration,
Biomaterials, Vol. 32, pp 1838-1847
Summary and Conclusions
1. Cardiac co-cultures can be seeded in different sequences and
cultured in peptides incorporated 3D alginate scaffolds.
2. CMs show a more elongated morphology when seeded after ECs.
3. In EC-FB co-cultures, cell numbers and viability remains stable.
4. Collagen and laminin production increases over time.
2012-50
B CA B
EC – FB co-culture
0
1000
2000
3000
4000
5000
6000
7000
8000
D3 D7 D10
Hoechst(FU)
DNA content
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
D3 D7 D10
Alamarblue(FU)/Hoechst(FU)
Viability relative to DNA content
0
2
4
6
8
10
12
D3 D7 D10
Collagen(ng)/Hoecst(FU)
Total collagen relative to DNA content
Figure 2: Analysis of EC-FB co-culture during 10 days. DNA
content (Hoechst, left), cell viability (Alamar blue, middle)
and total collagen (Sircol, right). Only total collagen results
show a significant difference (p = 0.04, one way ANOVA).
Laminin expression in EC – FB co-culture
Figure 3: Confocal imaging of
laminin expression.
Immunostaining for laminin
(red), and DAPI (blue). (A) 3
days post seeding. (B) 10 days
post seeding. Bar=50 µm
EC – HMEC-1 cell line
FB – rat neonatal cardiac fibroblasts
CM – rat neonatal cardiomyocytes
A
A B