||Peripheral blood mononuclear cells (PBMCs)
|Donor age at sampling
||1 million cells
||vapour phase nitrogen
Frequently Asked Questions
Do you have a protocol for plating Axol iPSC-Cardiomyocyte onto MEA plate ?
Yes, please follow the link here: https://www.axolbio.com/page/how-to-mea-atrial-and-ventricular-cardiomyocytes
What is the difference among the three ventricular cardiomyocyte products of different donor origin?
ax2502 is derived from a female 42 year-old donor's PBMCs and Sendai virus reprogramming is used for generating its parental iPSC line.
ax2505 is derived from CD34+ cord blood cells (male) that were reprogrammed using episomal vectors. These iPSCs have been used to generate ax0015 (iPSC-derived Neural Stem Cells), ax2115 (iPSC-derived Renal Proximal Tubular Cells) and ax0055 (iPSC-derived Sensory Neuron Progenitors).
ax2520 is derived from CD34+ cord blood cells (male) that were reprogrammed using episomal vectors under current good manufacturing practice (cGMP) conditions. From the master stock of cGMP iPSCs a non-GMP, research grade working bank of iPSCs was created. Axol has then differentiated these research grade iPSCs into ventricular cardiomyocytes, this means that these cells are ideal for pre-clinical research as the iPSCs from cGMP are available for future clinical work.
Please see publication below for information on this line of iPSCs:
Baghbaderani BA, Tian X, Neo BH, et al. cGMP-Manufactured Human Induced Pluripotent Stem Cells Are Available for Pre-clinical and Clinical Applications. Stem Cell Reports (2015)
Issues with cardiomyocyte aggregates after thawing
1. At the resuspension step, using P1000 and 1ml of plating medium to pipette up and down at least 5 times gently against the falcon tube which would partially disintegrate the aggregates;
2. In the counting step do not count the large aggregates and seed the culture vessel at a density of 50,000 cells/cm2 as instructed by the protocol;
3. The day after plating (Day 1), change Plating Medium to Cardiomocyte Maintenance Medium which would remove debris and cells that didn't attach (including large aggregates).
4. Using the recommended seeding density we have achieved over 70% confluence culture after plating.
Can I expand the ventricular cardiomyocytes?
No, you may passage within 3 days of plating to change culture vessel, but this is not recommended.
Can I freeze and thaw cardiomyocytes after culturing?
No, because the extra freeze/thaw cycle would effect the viability of the cells.