Cultured murine bone marrow derived mesenchymal stem cells (BMSC) when grown along with cardiogel derived from mouse cardiac fibroblast, exhibited increased cell proliferation and differentiation and enhanced survival under oxidative stress induced by the exposure of H2O2 in vitro (similar to in vivo ischemia like condition). Adhesion of BMSC to the cardiogel occurred at a faster rate when compared to the cells grown on normal surface. BMSC attached to cardiogel showed an increased resistance to proteolytic (enzymatic) disassociation. This is the first report on an attempt to use an in house biomaterial for the growth of BMSC that led to their heightened resistance towards oxidative stress. These studies support that cardiogel is an efficient biodegradable three-dimensional extracellular matrix which supports better growth of BMSC and can be used as a scaffold for stem cell delivery, with potential therapeutic applications in cardiac tissue regeneration.

Rama Shanker Verma

Department Of Biotechnology

Biomaterial

Hydrogen peroxide

animal cell

article

binding affinity

Biodegradability

CARDIOGEL

Cell adhesion

cell differentiation

cell growth

cell proliferation

cell survival

controlled study

extracellular matrix

mesenchymal stem cell

mouse

MYOFIBROBLAST

nonhuman

oxidative stress

protein degradation

Fulltext

IIT Madras is a public technical and research university located in Chennai, Tamil Nadu. Founded in 1959, it is recognised as an Institute of National Importance.

IIT Madras has been ranked as the top engineering institute in India for four years in a row by the National Institutional Ranking Framework of the MHRD

It currently offers undergraduate, postgraduate and research degrees across 16 disciplines in Engineering, Sciences, Humanities and Management. About 596 faculty belonging to science and engineering departments and centres of the Institute are engaged in teaching, research and industrial consultancy.

IIT Madras

European Cells and Materials

Mesenchymal stem cells (MSC), because of their multipotency and ease of purification and amplification, are an ideal stem cell source for cell therapies. Bone-marrow-derived stem cells (BMSC) can be used to develop MSC-like immortalized cell lines with large proliferation and differentiation potentialities. Their immortalized status prevents the maintenance of MSC function and characters; this can be negated by modifying the isolation and maintenance protocol. Adult murine BMSC were isolated and maintained in media without additional growth factors together with passage-dependent reseeding following trypsinization. Cells maintained over 25 passages were considered as putative cell lines and characterized. The phenotypic and genotypic characteristics and multilineage differentiation potential of the cells were assessed by morphological, phenotypic, and molecular assays at various passages. The putative BMSC cell lines showed the characteristics of MSC and were able to maintain these characteristics, even after immortalization. The phenotypic data demonstrated difference among two cell lines; this was further validated by the difference in their multilineage differentiation potential following specific induction. More importantly, no changes were observed in the genotypic level in comparison with control cells, even after more than 50 passages. Our protocol thus advances the isolation and maintenance of BMSC and the development of putative BMSC cell lines that maintain characteristics of MSC, including multilineage differentiation potential, after more than 40 passages. © 2012 Springer-Verlag.

Cell and Tissue Research

Generation of mesenchymal stem cell lines from murine bone marrow

The lack of an established protocol for scanning electron microscopy (SEM) studies on stem cells differentiating into adipogenic lineage led us to develop a protocol for the preparation of differentiated adult bone marrow-derived mesenchymal stem cells (BMSC) for SEM. This protocol describes the procedure to maintain and preserve the structural organization of cellular components following differentiation, for morphological and physical characterization. The fixation of the differentiated cells was followed by dehydration using methanol, and vacuum desiccation before microscopy. The use of longer chain alcohols as dehydrating agents was avoided in our method to reduce the dissolution of lipid deposits in cells, thus allowing the maintenance of their structural integrity. The time period for the processing of samples was reduced by avoiding the osmium tetroxide postfixation and critical point drying. Thus, this protocol helps in determining the potential, fate, and degree of stem cell differentiation. This may be useful for SEM analysis of differentiated cells, especially those grown on various scaffolds. © 2011 Elsevier Inc. All rights reserved.

Analytical Biochemistry

Scanning electron microscopy preparation protocol for differentiated stem cells

Cardiogel supports adhesion, proliferation and differentiation of stem cells with increased oxidative stress protection

Journal	European Cells and Materials
Publisher	AO Research Institute Davos
ISSN	14732262
Open Access	Yes