Restoring mitochondrial function promotes hematopoietic reconstitution from cord blood following cryopreservation-related functional decline
Yaojin Huang, … , Yingchi Zhang, Tao Cheng
Yaojin Huang, … , Yingchi Zhang, Tao Cheng
Published March 4, 2025
Citation Information: J Clin Invest. 2025;135(9):e183607. https://doi.org/10.1172/JCI183607.
View: Text | PDF
Research Article Hematology

Restoring mitochondrial function promotes hematopoietic reconstitution from cord blood following cryopreservation-related functional decline

Abstract

Umbilical cord blood (UCB) plays substantial roles in hematopoietic stem cell (HSC) transplantation and regenerative medicine. UCB is usually cryopreserved for years before use. It remains unclear whether and how cryopreservation affects UCB function. We constructed a single-cell transcriptomics profile of CD34+ hematopoietic stem and progenitor cells (HSPCs) and mononuclear cells (MNCs) from fresh and cryopreserved UCB stored for 1, 5, 10, and 19 years. Compared with fresh UCB, cryopreserved HSCs and multipotent progenitors (MPPs) exhibited more active cell-cycle and lower expression levels of HSC and multipotent progenitor signature genes. Hematopoietic reconstitution of cryopreserved HSPCs gradually decreased during the first 5 years but stabilized thereafter, aligning with the negative correlation between clinical neutrophil engraftment and cryopreservation duration of UCB. Cryopreserved HSPCs also showed reduced megakaryocyte generation. In contrast, cryopreserved NK cells and T cells maintained a capacity for cytokine production and cytotoxicity comparable to that of fresh cells. Mechanistically, cryopreserved HSPCs exhibited elevated ROS, reduced ATP synthesis, and abnormal mitochondrial distribution, which collectively led to attenuated hematopoietic reconstitution. These effects could be ameliorated by sulforaphane (SF). Together, we elucidate the negative effect of cryopreservation on UCB HSPCs and identify SF as a mitigation strategy, broadening the temporal window and scope for clinical applications of cryopreserved UCB.

Authors

Yaojin Huang, Xiaowei Xie, Mengyao Liu, Yawen Zhang, Junye Yang, Wenling Yang, Yu Hu, Saibing Qi, Yahui Feng, Guojun Liu, Shihong Lu, Xuemei Peng, Jinhui Ye, Shihui Ma, Jiali Sun, Lu Wang, Linping Hu, Lin Wang, Xiaofan Zhu, Hui Cheng, Zimin Sun, Junren Chen, Fang Dong, Yingchi Zhang, Tao Cheng

×

Figure 2

Substantial transcriptomic variations occur in HSPCs after cryopreservation.

Options: View larger image (or click on image) Download as PowerPoint
Substantial transcriptomic variations occur in HSPCs after cryopreservat...
(A) The number of DEGs in HSPCs between 2 consecutive periods. (B) Bubble plot showing the enriched GO terms of upregulated genes in HSPCs from cryopreserved UCB (cryopreserved UCB vs. fresh UCB; UCB from different cryopreservation years was analyzed as a whole). Pos. reg., positive regulation. (C) Percentages of HSCs, MPP1s, MPP2s, and Mitohi HSCs/MPPs in different cell-cycle phases for fresh and cryopreserved UCB stored for 1, 5, 10, and 19 years. For each cluster, the proportions of cells in the G2M phase at 1 year, 5 years, 10 years, and 19 years were respectively compared with that of fresh UCB using the χ2 test; data indicate the mean ± SD (see the Supporting Data Values file). (D) Relative expression of stemness signature genes in HSCs, MPP1s, MPP2s, and Mitohi HSCs/MPPs from fresh and cryopreserved UCB stored for 1, 5, 10, and 19 years. **P ≤ 0.01 and ****P ≤ 0.0001, by Wilcoxon test. The stemness signature genes used for the analysis are listed in Supplemental Data File 2. (E) Potential of Mitohi HSC/MPP differentiation toward MLP, GMP, and MEP lineages for fresh and cryopreserved UCB. UCB samples from different cryopreservation years were analyzed as a whole.