Blood donor-derived buffy coat to produce platelets in vitro

Vox Sang. 2020 Jan;115(1):94-102. doi: 10.1111/vox.12863. Epub 2019 Nov 10.

Abstract

Background and objectives: Platelet transfusion is a standard medical therapy used to treat several bleeding disorders. However, a critical drawback is the dependency on donor-derived platelets, which leads to concerns like insufficient availability and immunological complications. In vitro platelet production from hematopoietic progenitor cells (CD34) may represent a reasonable solution.

Materials and methods: CD34+ cells were isolated from either buffy coat or peripheral blood and compared in terms of platelet production in vitro. The number and the quality of magnetically isolated CD34+ cells and their capability to differentiate into mature megakaryocytes were investigated using flow cytometry. Additionally, the functionality of megakaryocytes in term of in vitro platelet production was tested.

Results: Similar purity and quantity of CD34+ cells was found after their isolation from both cell sources. In contrast, after 6 days of culture, enhanced number of CD34+ cells isolated from buffy coat compared with peripheral blood was observed (5·3 x 106 vs. 3·0 x 106, respectively). Interestingly, despite a comparable nuclear maturation phenotype, the yield of platelets released from buffy coat-derived megakaryocytes was significantly higher than from peripheral blood cells (platelet yield pro MK: 7·2 vs. 2·7, respectively). Importantly, platelets produced from buffy coat-derived cells could be activated by agonists.

Conclusion: Haematopoietic progenitor cells isolated from buffy coat have increased yield of platelets released from mature megakaryocytes and enhanced in vitro functionality, compared with peripheral blood-derived cells. Our study, suggests that buffy coat, obtained during blood donation processing, might be a promising source of megakaryocytes for in vitro platelet production.

Keywords: hematopoietic stem cells; megakaryocytes; platelet generation; thrombocytopenia.

MeSH terms

  • Blood Buffy Coat / cytology*
  • Blood Donors*
  • Blood Platelets*
  • Flow Cytometry
  • Hematopoiesis
  • Hematopoietic Stem Cells / physiology*
  • Humans
  • Megakaryocytes*