I would be using 2 layered cell factory (1272cm2) for Mesenchymal stem cell culture.
Source - P1 Umbilical cord Mesenchymal Stem Cell.
If it is feasible to culture and arrive at the magical number (I100 Million mesenchymal stem cell ). what would be the ideal media to use, Supplement ( FBS % and bFGF concentration ) and Seeding density ?
Hi Balasubramani,
I believe it'll be possible to harvest 100 million MSCs. You can calculate the number you need to seed it using the formula PD=log(harvest-seeded)/log2. you can also always pool the clonal MSC and count from P1.
after the culture is made you need to count the doubling time (d.t.) for each passage (divide the time in culture by the PD).
you need also to consider that the growth will be slower during the time, it will be slower in day 15 than day 5 of your culture.
you can refer to this question below to deepen your understanding.
regards.
https://www.researchgate.net/post/What_is_the_population_doubling_time_of_mesenchymal_stem_cells_derived_from_human_bone_marrow_derived
Dear Alex,
Here are my results from 4 different 2 layered cell factory !!
Cell Factory #1 - 77 million
Cell Factory #2 - 63 million
Cell Factory #3 - 56 million
Cell Factory #4 - 43 million.
70-80% confluent flask (any type) will have ~20,000 - 30,000 cells/sq.cm. if you calculate based on this a two layer cell factory (~1260 sq cm) should not have more than 40 million msc at 70-80% confluence. and the recovery of cells from the flask by trypsinization will not be 100% you will still have some cells left in the cell factory. recovery will be around 80-90%. so the final yield from a two layer cell factory will be 30-35 million msc.
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