Reducing Mitomycin-C-

induced ROS levels in

z mouse feeder cells
improves induced
pluripotent stem cell
colony growth
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TERMINOLOGY:

Induced Pluripotent Stem Cells (iPSC) – are somatic cells that

have been reprogrammed to behave in an embryonic pluripotent state and is
used for therapeutic purposes.

Feeder Cells – growth arrested cells that are still biologically active. They
serve as support to ensure the growth of primary cells or function as cytokines
wherein it signals to regulate the growth and activities done by the immune
system

Feeder-dependent Culture – is a process of stem cell culture used to

retain stem cells that require “feeder cell” co-culture, such as fibroblasts, to
facilitate the pluripotency and proliferative ability of these stem cells.

Cell viability – a measure of the number of live, healthy cells within a

population.
 Embryonic Stem Cells (ESCs) -derived from early embryos and are capable of indefinite
z
self renewal while maintaining the potential development of cells; ESCs are pluripotent,
meaning they can form tissues from the three primary germ layers. 


Mouse Embryonic Fibroblast- a type of fibroblast that is taken from a mouse embryo .


Mitomycin-C (MMC) - are derived from Streptomyces caespitosus  It is an alkylating agent

that inhibits DNA synthesis and cell proliferation. [A method used for inactivating Feeder Cells]


Gamma-irradiation - is a physical or chemical means of sterilization, which kills bacteria by

destroying bacterial DNA. Thus, suppressing the bacterial division. It is a process that
incorporates the use of Cobalt 60. Moreover, it is used for decontamination and modification.
[A method used for inactivating Feeder Cells]


MTT Assay - used for the evaluation of a cell’s metabolic activity. It is used as a measure of
cell viability, proliferation and cytotoxicity. 
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Introduction
Research
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Question/s 
 How does mitomycin-C relate to reactive oxygen species?

 How does the length of exposure to mitomycin-C affect feeder cells?

 How do stress in feeder cells affect iPSC growth during coculture?

Research Objectives
 To show how to effectively arrest MEFs without inducing oxidative stress by
optimization of MMC exposure time. 

 To determine the effect of mytomycin-C (MMC) and its length of exposure.

 To determine whether stress in feeder cells (MEF) indirectly affects iPSC growth during
coculture.  

 To establish budget-efficient culture strategies for adapting stem cell-based

applications.