A PhD student leaving for vacation has asked an undergraduate student to perform daily media changes for his iPSCs while he is gone. The culture is happening in 12-well plates where a volume of 2 mL is optimal. On Saturday, when changing the media, the undergraduate decides to add 4 ml of media to the dishes (instead of 2 ml) because he wants to skip lab and watch the Super Bowl on Sunday. He decides to add twice the volume of media (4 mL) to tide the cells over till Monday. However, when the graduate student returns on Monday, he finds that some of his cells have died. Your job is to determine whether the cells died due to a lack of oxygen. For the calculations that follow, diffusion and reaction occurs only in one direction. Also, assume that reaction only occurs at the cell-media interface. Use Michaelis-Menten type kinetics for oxygen uptake. You may use the following information: Ps = 150 mmHg (ambient oxygen tension) K = 1.19 nmol / mL / mmHg (solubility of oxygen in medium) D = 2 ×10−5cm2/s (diffusivity of oxygen through medium at 37 °C) Km = 0.5 mmHg (partial pressure at which O2uptake rate is half maximal) Vmax = 3 ×10−5nmol / s / cell (maximum oxygen uptake rate) Cell viability is compromised at an oxygen tension of Pδ = 0.5 mmHg or below. The flux of oxygen due to diffusion through the medium is given by D*(K(Ps −Pδ))/δ where δ is the height of the medium. Part 2.2 If a 6-well dish (9.6 cm2) was used instead of a 12-well plate, and if cells were seeded at the same cell density (10,000 cells/cm2) as before, what would be the critical volume of media to maintain cell viability? Would the increase in media volume be directly proportional to this increase in dish surface area?

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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A PhD student leaving for vacation has asked an undergraduate student to perform daily media changes for
his iPSCs while he is gone. The culture is happening in 12-well plates where a volume of 2 mL is optimal. On
Saturday, when changing the media, the undergraduate decides to add 4 ml of media to the dishes (instead
of 2 ml) because he wants to skip lab and watch the Super Bowl on Sunday. He decides to add twice the
volume of media (4 mL) to tide the cells over till Monday. However, when the graduate student returns on
Monday, he finds that some of his cells have died. Your job is to determine whether the cells died due to a
lack of oxygen. For the calculations that follow, diffusion and reaction occurs only in one direction. Also,
assume that reaction only occurs at the cell-media interface. Use Michaelis-Menten type kinetics for oxygen
uptake. You may use the following information:
Ps = 150 mmHg (ambient oxygen tension)
K = 1.19 nmol / mL / mmHg (solubility of oxygen in medium)
D = 2 ×10−5cm2/s (diffusivity of oxygen through medium at 37 °C)
Km = 0.5 mmHg (partial pressure at which O2uptake rate is half maximal)
Vmax = 3 ×10−5nmol / s / cell (maximum oxygen uptake rate)
Cell viability is compromised at an oxygen tension of Pδ = 0.5 mmHg or below. The flux of oxygen due to
diffusion through the medium is given by
D*(K(Ps −Pδ))/δ
where δ is the height of the medium.

Part 2.2
If a 6-well dish (9.6 cm2) was used instead of a 12-well plate, and if cells were seeded at the same cell density
(10,000 cells/cm2) as before, what would be the critical volume of media to maintain cell viability? Would
the increase in media volume be directly proportional to this increase in dish surface area?

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