Elsevier

Transplantation Proceedings

Volume 39, Issue 7, September 2007, Pages 2434-2437
Transplantation Proceedings

Bone marrow and stem cell
Umbilical Cord Blood-Derived Stem Cells Spontaneously Express Cardiomyogenic Traits

https://doi.org/10.1016/j.transproceed.2007.06.016Get rights and content

Abstract

Background

Umbilical cord blood (UCB) has been widely used for hematopoietic stem cell transplantation. The UCB-derived stem cells (UCBSCs) have been proposed as an alternative to bone marrow (BM)–derived mesenchymal stem cells (MSCs) for cardiac cell-based therapy. Herein we studied whether UCBSCs spontaneously exhibit cardiac-specific markers in vitro.

Methods

Human UCBSCs were isolated, expanded, and phenotyped by flow cytometry, quantitative RT-PCR, and immunofluorescence. Cell pluripotency and proliferation were also assessed by adipogenic and osteogenic media and in growth assays.

Results

Among 25 analyzed UCB, 16% of cases afforded primary culture satisfactory generation of UCBSCs. Duplication time (Td) of cultures was 2.16 ± 0.06 days. The cells were strongly positive for CD105 (18.5 ± 0.14), CD44 (27 ± 2.8), CD166 (13 ± 9), CD29 (59 ± 9.4), CD90 (60 ± 11) and consistently negative for CD117 (1.2 ± 0.1), CD106 (1.1 ± 0), CD34 (1.2 ± 0.2), CD14 (1 ± 0), and CD45 (1 ± 0), consistent with a mesenchymal lineage. Adipogenesis and osteogenesis of cells resulted in low accumulation of intracellular lipid droplets and high deposition of calcium. The UCBSCs showed gene transcripts for α-actinin, connexin (Cx)-43, SERCA-2, and stromal cell-derived factor (SDF)-1α. At the protein level, the cells abundantly expressed α-actinin, Cx-43, SERCA-2 and SDF-1α. In contrast, these cells did not express the cardiac transcription factors GATA-4, Tbx5, and Nkx2.5, nor the sarcomeric proteins β-myosin heavy chain (β-MyHC) or cardiac troponin I (cTnI).

Conclusions

Human UCBSCs may represent an alternative source of stem cells for myocardial-cell replacement. These cells can be highly expanded. They spontaneously express proteins of paramount importance for cardiovascular regeneration, such as Cx-43, SERCA-2, and SDF-1α.

Section snippets

Collection of UCB, Isolation, and Cell Culture of UCBSCs

A total of 25 UCB samples (60–100 mL) were collected from the umbilical cord vein and processed within 12 hours after extraction. Cells were isolated according to modifications of a previously reported method.2 Blood cells clarified by previous centrifugation were resuspended in 30 mL calcium- and magnesium-free phosphate-buffered saline (PBS) (Invitrogen). The cell suspension layered over 1.077 g/mL Lymphoprep (Nycomed) was centrifuged at 400g for 30 minutes. Mononuclear cells recovered by

Results

The study included 25 UCB samples that were analyzed for their capacity to generate primary cultures of UCBSCs. Four primary cultures of UCBSCs were satisfactorily generated (16%). After selection and plating of mononuclear cells, a few elongated fibroblast-like cells attached to the plastic culture dishes were present within 4 weeks and expanded rapidly (Fig 1A, B). Primary cultures were maintained uninterruptedly over 4 months and 10 passages.

Induction of UCBSCs with adipogenic and osteogenic

Discussion

This study demonstrated that UCBSCs exhibited some cardiomyogenic lineage markers during in vitro culture in the absence of cardiac induction factors.

The advent of stem cell biology has provided a good option to test UCBSC’s properties and therapeutic application in myocardial regeneration. Thus, the foundation for the view of the heart as a terminally differentiated post-mitotic organ, which had profoundly conditioned basic and clinical research in cardiology, has been certainly rejected.8

Acknowledgments

The authors gratefully appreciate Berta Raposo for technical assistance in flow cytometry.

References (11)

There are more references available in the full text version of this article.

Cited by (46)

  • Stem Cell Therapy in Single-Ventricle Physiology: Recent Progress and Future Directions

    2021, Seminars in Thoracic and Cardiovascular Surgery: Pediatric Cardiac Surgery Annual
    Citation Excerpt :

    In fact, UCB-MSCs have demonstrated superior ex vivo expansion.16 Early in vitro expansion and characterization established expression of proteins important for cardiac regeneration (connexin-43, sarcoendoplasmic reticulum calcium transport ATPase 2, and stromal cell-derived factor-1α [SDF-1α).15 In addition to the classic MSC lineages, UCB-MSCs are capable of differentiating into many cell types including hepatocyte-like cells, neuroglial-like cells, respiratory epithelial cells, and cardiomyocytes.17-19

  • Umbilical cord blood-derived mesenchymal stem cells: New therapeutic weapons for idiopathic dilated cardiomyopathy?

    2014, International Journal of Cardiology
    Citation Excerpt :

    The most marked limitation in the use of UCB is its low progenitor cell concentration, albeit transplantation of double partially HLA-matched UCB units is recognized as a simple approach for overcoming cell dose limitation [73]. As mentioned above UCB also contains MSCs [74], which play key roles in the regulation of blood vessel formation [75]. UCBMSCs were initially characterized as spontaneously exhibiting cardiomyogenic traits, such as abundant expression of α-actinin, connexin-43, and SERCA-2 [74].

View all citing articles on Scopus

This work was supported by grants from Ministerio de Educación y Ciencia (SAF 2004-08044-C03-01). We also appreciate support from BMS, Fundación Daniel Bravo Andreu, and Fundación Roviralta.

1

C. Prat-Vidal and S. Roura contributed equally to this work.

View full text