Improving Human Induced Pluripotent Stem Cell-Derived Megakaryocyte Differentiation and Platelet Production

International Journal of Molecular Sciences, 2021 · DOI: 10.3390/ijms22158224 · Published: July 30, 2021

Immunology Regenerative Medicine & Stem Cells

Simple Explanation

The study focuses on improving the production of platelets from human induced pluripotent stem cells (hiPSCs). The researchers modified a protocol to enhance the differentiation of hiPSCs into megakaryocytes (MKs), which are cells that produce platelets. The modified protocol involves a mesoderm induction step before hemogenic endothelial cell (HEC) differentiation, which leads to increased production of MK progenitors (MKPs) and platelets. The study also examined the impact of different media compositions on MK maturation. The regenerative properties of hiPSC-derived platelets were assessed by comparing the expression of angiogenesis-related proteins to those of adult donor-derived platelets. The study found that hiPSC-derived platelets have a unique protein expression profile and may offer a sustainable source of pro-angiogenic factors for regenerative medicine.

Study Duration

Not specified

Participants

Healthy volunteers and blood donors

Evidence Level

Not specified

Key Findings

1
A modified protocol including activin A and CHIR99021 significantly increased HEC differentiation, resulting in improved MK production.
2
The modified protocol resulted in a five-fold increase in platelet production per hiPSC compared to standard conditions (235 ± 84 vs. 51 ± 15).
3
hiPSC-derived platelets showed comparable expression of 19 out of 24 angiogenesis-related proteins compared to adult donor-derived platelets, with some proteins being expressed at higher or lower levels.

Research Summary

This study demonstrates that distinct preceding induction by activin A, BMP-4, CHIR, and VEGF in lipid-enriched APEL medium resulted in a signiﬁcant increase of hiPSC-derived platelet production using an otherwise unaltered three-stage standard protocol. From a mechanistic point of view, the major effect observed after an initial induction step appeared to be a more robust HEC formation (stage I). We found nine-fold higher MKP proportions already at day 9 (stage II) of the improved protocol compared to standard cultures as illustrated in tSNE plots. The rich pro-angiogenic proteome of hiPSC-derived platelets did not differ irrespective of initial induction. Signiﬁcant differences between adult donor-derived and hiPSC-derived platelet proteomes may at least in part be donor-dependent requiring future extended analysis.

Practical Implications

Improved Platelet Production

The modified protocol offers a more efficient method for generating platelets from hiPSCs, potentially addressing the increasing need for blood products.

Regenerative Medicine Applications

hiPSC-derived platelets may serve as a sustainable source of pro-angiogenic factors for regenerative medicine therapies, replacing fetal bovine serum.

Clinical Transfusions

The ability to manufacture platelets from hiPSCs could provide clinical platelet units for patients with alloimmunization against frequent antigens, who currently lack suitable donors.

Study Limitations

1
Functional assays are partly limited by the low numbers of platelets produced in vitro.
2
We realized a certain level of variability in our 2D culture protocol, which leaves space for further improvement.
3
Significant differences between adult donor-derived and hiPSC-derived platelet proteomes may at least in part be donor-dependent requiring future extended analysis.

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