4 Notes
1. The range and amounts of cell culture maintenance consumables should be adapted to particular experimental needs, such
as the type of MK differentiation protocol applied, and desired
volumes.
2. Water for cell culture applications may be required for reconstitution of certain cytokines and preparation of other solutions
according to specifications.
3. TrypLE™ Express Enzyme may be required for detachment of
iPSC cells during cell culture maintenance. It is recommended
to use TrypLE™ Express Enzyme instead of conventional
trypsin due to a milder action on the cells.
4. Use medium appropriate for differentiation of iPSCs into myeloid lineage. Choice of medium depends on particular differentiation protocol and experimental requirements selected for
generation of iPSC-derived MKs [1, 10, 14, 21].
5. Use a set of cytokines and growth factors necessary for differentiation of iPSCs into MKs at appropriate time points of
differentiation procedure. Essential cytokines and growth factors include, but not limited by, vascular endothelial growth
factor (VEGF), thrombopoietin (TPO), and stem cell factor
(SCF). Selection of cytokines and growth factors, as well as
their concentrations, depends on particular differentiation protocol and experimental requirements chosen for generation of
iPSC-derived MKs [1, 10, 14, 21].
6. Select iPSC line capable of differentiation into MKs. Different
rates of differentiation outcome may depend on iPSC origin
and levels of pluripotency gene expression [1, 21]. In addition
to iPSCs, it is also possible to differentiate certain other cell
types into MKs [4, 6–9].
7. A programmable freezer may be also applied for optimizing
cooling rates, which may be required for CPAs alternative
to DMSO.
8. CPAs alternative to DMSO may be tested for cryopreservation
activity. However, concentrations and cooling rates have to be
optimized accordingly.
9. Selection of cytokines and growth factors, as well as their concentrations, depends on particular differentiation protocol and
experimental requirements chosen for generation of iPSCderived MKs [1, 10, 14, 21]. Generally, iPSCs may be differentiated as a monolayer culture or in bioreactors [10, 14]. In
addition to iPSCs, it is also possible to differentiate certain
other cell types such as hematopoietic CD34+ cells or MSCs
into MKs [4, 6–9].
552
Denys Pogozhykh et al.
1. The range and amounts of cell culture maintenance consumables should be adapted to particular experimental needs, such
as the type of MK differentiation protocol applied, and desired
volumes.
2. Water for cell culture applications may be required for reconstitution of certain cytokines and preparation of other solutions
according to specifications.
3. TrypLE™ Express Enzyme may be required for detachment of
iPSC cells during cell culture maintenance. It is recommended
to use TrypLE™ Express Enzyme instead of conventional
trypsin due to a milder action on the cells.
4. Use medium appropriate for differentiation of iPSCs into myeloid lineage. Choice of medium depends on particular differentiation protocol and experimental requirements selected for
generation of iPSC-derived MKs [1, 10, 14, 21].
5. Use a set of cytokines and growth factors necessary for differentiation of iPSCs into MKs at appropriate time points of
differentiation procedure. Essential cytokines and growth factors include, but not limited by, vascular endothelial growth
factor (VEGF), thrombopoietin (TPO), and stem cell factor
(SCF). Selection of cytokines and growth factors, as well as
their concentrations, depends on particular differentiation protocol and experimental requirements chosen for generation of
iPSC-derived MKs [1, 10, 14, 21].
6. Select iPSC line capable of differentiation into MKs. Different
rates of differentiation outcome may depend on iPSC origin
and levels of pluripotency gene expression [1, 21]. In addition
to iPSCs, it is also possible to differentiate certain other cell
types into MKs [4, 6–9].
7. A programmable freezer may be also applied for optimizing
cooling rates, which may be required for CPAs alternative
to DMSO.
8. CPAs alternative to DMSO may be tested for cryopreservation
activity. However, concentrations and cooling rates have to be
optimized accordingly.
9. Selection of cytokines and growth factors, as well as their concentrations, depends on particular differentiation protocol and
experimental requirements chosen for generation of iPSCderived MKs [1, 10, 14, 21]. Generally, iPSCs may be differentiated as a monolayer culture or in bioreactors [10, 14]. In
addition to iPSCs, it is also possible to differentiate certain
other cell types such as hematopoietic CD34+ cells or MSCs
into MKs [4, 6–9].
552
Denys Pogozhykh et al.