Functional properties of cells obtained from human cord blood CD34 + stem cells and mouse cardiac myocytes in coculture

Alessia Orlandi, Francesca Pagani, Daniele Avitabile, Giuseppina Bonanno, Giovanni Scambia, Elisa Vigna, Francesca Grassi, Fabrizio Eusebi, Sergio Fucile, Maurizio Pesce, Maurizio C. Capogrossi

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Prior in vitro studies suggested that different types of hematopoietic stem cells may differentiate into cardiomyocytes. The present work examined whether human CD34+ cells from the human umbilical cord blood (hUCB), cocultured with neonatal mouse cardiomyocytes, acquire the functional properties of myocardial cells and express human cardiac genes. hUCB CD34+ cells were cocultured onto cardiomyocytes following an infection with a lentivirus-encoding enhanced green fluorescent protein (EGFP). After 7 days, mononucleated EGFP+ cells were tested for their electrophysiological features by patch clamp and for cytosolic [Ca2+] ([Ca 2+]i) homeostasis by [Ca2+]i imaging of X-rhod1-loaded cells. Human Nkx2.5 and GATA-4 expression was examined in cocultured cell populations by real-time RT-PCR. EGFP+ cells were connected to surrounding cells by gap junctions, acquired electrophysiological properties similar to those of cardiomyocytes, and showed action potential-associated [Ca2+]i transients. These cells also exhibited spontaneous sarcoplasmic reticulum [Ca2+]i oscillations and the associated membrane potential depolarization. However, RT-PCR of both cell populations showed no upregulation of human-specific cardiac genes. In conclusion, under our experimental conditions, hUCB CD34+ cells cocultured with murine cardiomyocytes formed cells that exhibited excitation-contraction coupling features similar to those of cardiomyocytes. However, the expression of human-specific cardiac genes was undetectable by RT-PCR.

Original languageEnglish
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume294
Issue number4
DOIs
Publication statusPublished - Apr 2008

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Coculture Techniques
Fetal Blood
Cardiac Myocytes
Stem Cells
Blood Cells
Genes
Excitation Contraction Coupling
Polymerase Chain Reaction
Lentivirus
Intercellular Junctions
Gap Junctions
Sarcoplasmic Reticulum
Hematopoietic Stem Cells
Membrane Potentials
Population
Action Potentials
Real-Time Polymerase Chain Reaction
Homeostasis
Up-Regulation
Infection

Keywords

  • Functional differentiation

ASJC Scopus subject areas

  • Physiology

Cite this

Functional properties of cells obtained from human cord blood CD34 + stem cells and mouse cardiac myocytes in coculture. / Orlandi, Alessia; Pagani, Francesca; Avitabile, Daniele; Bonanno, Giuseppina; Scambia, Giovanni; Vigna, Elisa; Grassi, Francesca; Eusebi, Fabrizio; Fucile, Sergio; Pesce, Maurizio; Capogrossi, Maurizio C.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 294, No. 4, 04.2008.

Research output: Contribution to journalArticle

Orlandi, Alessia ; Pagani, Francesca ; Avitabile, Daniele ; Bonanno, Giuseppina ; Scambia, Giovanni ; Vigna, Elisa ; Grassi, Francesca ; Eusebi, Fabrizio ; Fucile, Sergio ; Pesce, Maurizio ; Capogrossi, Maurizio C. / Functional properties of cells obtained from human cord blood CD34 + stem cells and mouse cardiac myocytes in coculture. In: American Journal of Physiology - Heart and Circulatory Physiology. 2008 ; Vol. 294, No. 4.
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