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Three-Dimension Co-culture of Hematopoietic Stem Cells and Differentiated Osteoblasts on Gallic Acid Grafted-Chitosan Scaffold as a Model of Hematopoietic Stem Cells Niche

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Abstract

The existing approaches of hematopoietic stem cells (HSCs) expansion in vitro were difficult to meet the needs of clinical application. While in vivo, HSCs efficiently self-renew in niche where they interact with three dimension extracellular matrix and stromal cells. Osteoblasts (OBs) are one of most significant stromal cells of HSCs niche. Here, we proposed a three-dimensional environment based on gallic acid grafted-chitosan (2c) scaffold and OBs differentiated from human umbilical cord mesenchymal stem cells (HUMSCs) to recapitulate the main components of HSCs niche. The results of alkaline phosphatase staining and alizarin red staining demonstrated that HUMSCs were successfully induced into OBs. The results showed that the expansions of CD34+cells, CD34+CD38− cells and CD34+CD38−CD45RA−CD49f+CD90+ cells (primitive hematopoietic stem cells, pHSCs) harvested from the biomimetic HSCs niche based on 2c scaffold and OBs (IV) group were larger than those harvested from other three culture groups. Importantly, it was found that the CD34+ cells harvested from IV group had better secondary expansion capability and colony forming potential, indicating better self-renewal ability. In addition, the biomimetic HSCs niche based on 2c scaffold and OBs protected HSCs apoptosis and promoted HSCs division. Taken together, the biomimetic HSCs niche based on 2c scaffold and OBs was an effective strategy for ex vivo expansion of HSCs in clinical scale.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China, 2018YFC1105800.

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  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Jin Wang, Minghao Xiong, Qihao Sun, Wen-Song Tan & Haibo Cai

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  1. Jin Wang
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  2. Minghao Xiong
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Contributions

Jin Wang contributed to the study conception and design. Material preparation, experiments, data collection and analysis were conducted by Jin Wang, Minghao Xiong and Qihao Sun. Flow cytometry analysis and Immunocytochemistry experiments were performed by Jin Wang. The first draft of the manuscript was written by Jin Wang and Haibo Cai, revised by Wen-song Tan and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Haibo Cai.

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Wang, J., Xiong, M., Sun, Q. et al. Three-Dimension Co-culture of Hematopoietic Stem Cells and Differentiated Osteoblasts on Gallic Acid Grafted-Chitosan Scaffold as a Model of Hematopoietic Stem Cells Niche. Stem Cell Rev and Rep 18, 1168–1180 (2022). https://doi.org/10.1007/s12015-021-10325-5

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