Rat
cardiomyocytes and bone marrow cells in coculture
Josef Skopalik1, Michal Pasek1, Eva Gabrielova2, Milan Richtarik3, Martin Modriansky2, Martin Klabusay1
1 Faculty of Medicine, Masaryk
University, Brno, Czech Republic
2 Faculty of Medicine, Palacky
University, Olomouc, Czech Republic
3 Faculty of Electrical Engineering, University of Technology, Brno, Czech Republic
Introduction: Adult bone marrow stem
cells have been used for treatment
of infarcted hearth muscle (Meluzin et
al., 2006 and others). These studies suggest that improvement over
conventional therapy can be achieved. However, the mechanisms by which the stem
cells might improve myocardial function are still poorly understood (three
hypothesis have beeen proposed so far - Klabusay and Skopalík, 2009). The
interaction between myocytes and bone marrow cells in vitro has been analyzed
in several groups (Beeres et al., 2005). It has been used only neonatal myocyte
yet. In our lab we repeated some of this experiment with neonatal myocytes. We
started this type of experiment with
adult rat myocyte too.
Material and methods: A) Rat
neonatal and adult cardiac myocytes were isolated (methods published by Chlopcikova et al., 2001 and Bébarová
at al.., 2005), the cells were then cultured in medium with fetal bovine serum.
Viability and several other parametres were analyzed over 3 days of the
culture. B)
Cocultivation experiments were started: myocytes were seeded in 24-well tissue-culture plates,
calcein-loaded bone marrow cells were added and interaction between myocytes
and marrow cells were analysed by fluorescence
microscope over 48 hours.
Results and conlusions: A) Neonatal myocytes have no problems with
viability and functionality over one week of culture. The amount of adult viable
myocytes decreased only very slowly over 3 days of culture. These types of
myocytes are suitable for 24 and 48 hours in
vitro “stem cell - myocyte cocultivation experiments“. B) First results
from cocultivation experiments suggested that bone marrow cells cocultivation with neonatal
myocytes lead to development of conductive connection between these two types
of cell (in agreement with published results). Stem cells cocultivation with
adult myocytes suggested creating of
strong adherence, but conductive
connection has not been detected yet. These results suggests that bone marrow cells
have not only paracrine effect during repair of affected myocardium.
References
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Matejovic P, Pasek M, Simurdova M and Simurda J. Effect of Ajmaline on Action
Potential and Ionic Currentsin Rat Ventricular. Gen. Physiol. Biophys, 24, 311—325, 2005
- Beeres S, Atsma
ED , der Laarse A. Human Adult Bone Marrow Mesenchymal Stem Cells Repair Experimental Conduction Block
in Rat Cardiomyocyte Cultures. JACC, 46,
10, 1943-52, 2005
- Chlopcikova S, Psotova J,
Miketova P. Model for the study of
morphological, biochemical characteristics of the hearth. Biomed. Papers 145(2), 49–55, 2001
- Klabusay M, Skopalík J, Meluzín J.
Minulost, prítomnost a budoucnost celularni terapie v kardiologii (in czech). Interni medicina pro praxi, in press,
2009