Research on establishing a protocol for isolation of CD34+ stem cells from umbilical cord blood at Vietnam National Institute of Maritime Medicine and Hai Phong Medical University Hospital, 2024 – 2025

Abstract

Objectives: To establish a protocol for isolating hematopoietic stem cells from umbilical cord blood (UCB) at the Vietnam National Institute of Maritime Medicine and Hai Phong Medical University Hospital in 2024, and to evaluate the quality of CD34+ stem cells obtained from UCB. Subjects and Methods: A prospective experimental study was conducted on 30 samples of CD34+ hematopoietic stem cells collected from the UCB of voluntary donors at the Vietnam National Institute of Maritime Medicine and Hai Phong Medical University Hospital between July 2024 and March 2025. The study protocol included donor screening, mononuclear cell (MNC) separation, CD34+ cell isolation using magnetic beads, culture in STEMMACS HSC-CFU gel-based medium, and colony morphology assessment by Giemsa staining. Results: The mean maternal age was 29.76 ± 5.24 years; mean neonatal birth weight was 3.31 ± 0.3 kg; and mean UCB volume was 75.37 ± 32.06 ml. The post-processing viability of MNCs and CD34+ cells was 95 ± 3% and 93 ± 4%, respectively. The recovery efficiency of MNCs was 75.27%. MNC counts showed a strong correlation with CD34+ counts (r = 0.6307; p < 0.001), with each unit yielding an average of 129 ± 44 × 10⁶ MNCs and 1.53 ± 0.9 × 10⁵ CD34+ cells. On average, each sample generated 127.53 ± 9.07 colonies, with burst-forming unit-erythroid (BFU-E) and colony-forming unit-granulocyte (CFU-G) predominating. No megakaryocytic colonies were detected. Conclusion: This study demonstrates the effectiveness of the established culture protocol and confirms that UCB is a potential source of hematopoietic stem cells for regenerative medicine and transplantation. Both MNCs and CD34+ cells exhibited high viability and robust multilineage differentiation potential.