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Mesenchymal Stem Cell Identification After Delayed Cord Clamping

  • Regenerative Medicine: Original Article
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A Correction to this article was published on 02 February 2023

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Abstract

We sought to determine the feasibility of identifying and quantifying mesenchymal stem cells (MSCs) from umbilical cord blood (UCB) after delayed cord clamping in preterm and term births. We obtained 3 mL of UCB at various gestational ages after delayed cord clamping. UCB separated by density gradient centrifugation within 4 h of delivery was passed through magnetic bead micro-columns to exclude the CD34 + cell population. The samples were incubated with fluorescent-tagged mesenchymal cell marker antibodies CD 29, CD44, CD73, CD105, and hematopoietic cell marker CD45. The cell populations were analyzed by flow cytometry. Viable cells were assessed with 7-aminoactinomycin-D. The results were expressed in median (minimum to maximum) MSCs and compared between preterm and term samples. A total of 12 UCB samples (32–40 weeks) were obtained, 10 of which demonstrated MSCs, accounting for 0.0174% (0–14.7%) of the viable UCB mononuclear cells. MSCs comprised 0.148% (0.0006–1.59%) and 0.116% (0–14.7%) of the viable UCB mononuclear cells in the term (n = 5), 38.4 ± 1.3 weeks, and preterm (n = 7) samples, 34.6 ± 1.1, respectively, p = 0.17. There was an overall median of 96 (0–39,574) MSCs. There was no difference in the median numbers of MSCs identified between term and preterm UCB samples, 3384 (23–6042) and 36 (0–39,574), respectively, p = 0.12. Mesenchymal stem cells were identified and quantified in 5 of 7 preterm and all 5 term UCB 3-mL samples obtained after delayed cord clamping.

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Acknowledgements

The authors wish to acknowledge Kent E. Vrana, PhD, for provision of laboratory assistance in the department of pharmacology. We thank Nate Sheaffer from Penn State College of Medicine’s Flow Cytometry Core (RRID:SCR_021134) for assistance with flow cytometry analysis.

Funding

Funding for the study was provided by the Division of Maternal–Fetal Medicine and the Department of Obstetrics and Gynecology, Penn State College of Medicine.

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Contributions

All authors contributed to study conception and design. Material preparations, data collection, and analysis were performed by Emily R. Smith, MD, Kevin P. Yeagle, MD, and Nurgul Salli, PhD. William M. Curtin, MD, assisted with analysis and display of data. The first draft of the manuscript was provided by Dr. Smith. Drs. Smith and Curtin revised and edited the final manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to William M. Curtin.

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The Human Subjects Protection Office of the institutional review board approval was obtained prior to initiation of the study (PRAMS031730EP).

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Informed consent was completed with patients prior to sample processing.

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The work was presented in poster format at the Society for Reproductive Investigation 68th Annual Scientific Meeting on July 8, 2021, in Boston, Massachusetts.

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Smith, E.R., Curtin, W.M., Yeagle, K.P. et al. Mesenchymal Stem Cell Identification After Delayed Cord Clamping. Reprod. Sci. 30, 1565–1571 (2023). https://doi.org/10.1007/s43032-022-01129-0

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