Cell barcoding reveals lineage-dependent outcomes in hiPSC cardiac differentiation
Sogu Sohn1, Daylin Morgan1, Cody Callahan1
1Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX 78712-0238, United States.
Insights
Tracking human induced pluripotent stem cell (hiPSC) clonal heterogeneity improves cardiac differentiation. Specific hiPSC lineages show distinct differentiation fates, impacting outcomes for cardiovascular disease treatments.
Area of Science:
- Stem Cell Biology
- Cardiovascular Research
- Developmental Biology
Background:
- Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show promise for cardiovascular disease treatment.
- Clinical translation is hindered by heterogeneous differentiation outcomes due to complex molecular interactions.
- Current methods lack adequate control over these interactions, leading to variable results.
Purpose of the Study:
- To investigate the role of clonal lineage-dependent responses (fate priming) in hiPSC cardiac differentiation.
- To determine if controlling hiPSC clonal heterogeneity can improve differentiation outcomes.
- To address the overlooked variable of hiPSC clonal heterogeneity in cardiac differentiation protocols.
Main Methods:
- Utilized the ClonMapper cell barcoding platform to label, track, and isolate distinct hiPSC lineages from a single cell line.
- Compared cardiac differentiation outcomes between isolated single-clone hiPSC populations and heterogeneous multi-clone hiPSC populations.
- Analyzed lineage-specific differentiation preferences towards cardiomyocytes (CMs) or non-cardiomyocytes.
Main Results:
- Demonstrated that specific hiPSC lineages exhibit preferential differentiation into hiPSC-CMs or non-CMs, indicating fate priming.
- Observed significant differences in cardiac differentiation outcomes between single-clone populations exhibiting fate priming and heterogeneous populations.
- Confirmed that hiPSC lineage identity influences the efficiency and outcome of cardiac differentiation.
Conclusions:
- hiPSC clonal heterogeneity is a critical factor influencing cardiac differentiation efficiency and outcome.
- Targeting and controlling hiPSC lineage identity can potentially optimize hiPSC-CM production for therapeutic applications.
- Understanding and leveraging fate priming in hiPSC cardiac differentiation is essential for advancing cardiovascular regenerative medicine.
Abstract:
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have potential applications in treating cardiovascular disease but are currently limited in their clinical translation. This can be attributed in large part to the complex molecular and cellular interactions that underly cardiac differentiation, with current differentiation approaches yielding heterogeneous outcomes due to inadequate understanding and control of these interactions. We hypothesize that clonal lineage-dependent responses to differentiation contribute to these heterogeneous outcomes, and as such cardiac differentiations can be improved by tracking and controlling for hiPSC clonal heterogeneity, a variable often overlooked in current differentiation approaches. "Fate priming," wherein clonal lineage identity determines differentiation fate, has been demonstrated in other stem cell differentiation pathways. We investigated fate priming in hiPSC cardiac differentiation using the ClonMapper cell barcoding platform to label, track, and isolate distinct hiPSC lineages from the same cell line. We show that certain hiPSC lineages preferentially differentiate into hiPSC-CMs or non-CMs. After isolating lineages with apparent fate priming, we found significant differences in cardiac differentiation outcomes between these single-clone populations and heterogeneous, multi-clone hiPSC populations. These findings indicate that lineage identity influences hiPSC cardiac differentiation outcomes.
