Other tissue/organ transplantation
Could the coculture of skeletal myoblasts and mesenchymal stem cells be a solution for postinfarction myocardial scar?

https://doi.org/10.1016/j.transproceed.2004.03.056Get rights and content

Abstract

Currently two lines of research have been proposed for treatment of heart failure in an attempt to address its main cause: skeletal myoblast (SM) transplants, which increase the contractile muscular mass, and mesenchymal stem cell (MSC) transplants, which increase neoangiogenisis. The objective of this study was to establish methods whereby cocultures of SM and MSC proliferate and expand, making possible the interaction of these cell types prior to their transplantation to the myocardium. Seeking to support the survival of these cells after myocardial transplantation and achieve subsequent functional improvement, SM and MSC from 10 rats were isolated and cultivated in DMEM medium supplemented with 15% fetal calf serum, 1% ATB, and growth factors. Following plating in variable proportions of satellite cells/mononuclear cells namely 2:1, 1:1, 1:2, morphological observations were made regarding cell survival, adhesion to substrate, and confluence. After 48 hours nonadherent cells were aspirated from the flasks, leaving the adherent cells, SM, and MSC. The better level of cell proliferation was observed with the proportion 2:1 cocultivated at a concentration of 5 × 105/mL for 14 days. The results were satisfactory; the cell production was up to 108, increasing the chances of transplant success after myocardial infarction. Transplants with this model are ongoing.

Section snippets

Methods

The isolation of SM from sacrificed neonatal rats used the enzymatic dissociation technique reported by Delaporte et al,9 employing muscular fragments from the four limbs. The isolation of MSC of BM was performed by two different harvesting techniques; a puncture-aspiration and a sacrifice of animal. The collected material was processed using a Ficoll-Hypaque density gradient (d = 1.077) according to Boyüm.10 The isolated cells were plated as described below. Assays were performed in 25-cm2

Results

The 2:1 ratio of satellite cells/stem cell was chosen because upon morphological observation the vials were more confluent in this proportion. Ten cocultures were performed, but only four groups (I, II, VII, and IX) were transplanted (Table 1, Table 2).

Discussion

It is concluded that cultivation of two distinct cell groups simultaneously may optimize the results already obtained from the transplantation of individually cultivated SM and MSC. We have demonstrated the possibility of coinjection of cultured cells preceeded by cocultivation may facilitate autologous transplantation, and transplants with this model are ongoing.

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  • Immunophenotypic Expression by Flow Cytometric Analysis of Cocultured Skeletal Muscle and Bone Marrow Mesenchymal Stem Cells for Therapy Into Myocardium

    2008, Transplantation Proceedings
    Citation Excerpt :

    Collection of skeletal muscle was obtained by biopsy of the posterior tibiae muscle with SMC isolations performed by enzymatic dissociation in accordance with Delaporte et al.12 Collection of MonoSC was obtained by puncture aspiration of bone marrow from the iliac crest with isolation by Ficoll-Hypaque (d = 1.077) density gradient in accordance with Boyüm.13 Coculture of SMC and BMMeSC was established as described by Carvalho et al.14 All cultures started with the same cell concentration (5 × 105/mL) were maintained in an Incubator with 5% CO2 at 37°C in Dulbecco's Modified Eagle Medium (DMEM; Gibco, United States) supplemented with 15% fetal calf serum (FCS; Gibco, United States), 10 μg/mL of insulin growth factor type I (Sigma, St Louis, Mo, United States), 1% antibiotic (100 μg/mL streptomycin–100 μg/mL penicillin), and 10−6 mol/L dexamethasone), which was changed every 48 hours for 21 days. The cell count was performed in a hemocytometer.

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