A Systematic Approach to Media Development for Efficient Expansion of Neuroepithelial Stem Cells in a GMP Setting
(2024) KIMM01 20241Educational programmes, LTH
Department of Immunotechnology
- Abstract
- Neuroepithelial stem (NES) cells derived from induced pluripotent stem cells (iPSCs), hold promise as a novel cell therapy for treating various neurological disorders such as, neurodegenerative diseases, spinal cord injury, and stroke. NES cells’ therapeutic potential lies in their ability to differentiate into multiple cell types and restore neuronal function. However, in order for this therapeutic potential to be exploited several modifications must be made. One of the important aspects for upscaling production is that the cell culture media must adhere to good manufacturing practice (GMP) guidelines. Currently existing media is supplemented with a complex of nutrients called “B27”, which has a composition derived from non-human origin... (More)
- Neuroepithelial stem (NES) cells derived from induced pluripotent stem cells (iPSCs), hold promise as a novel cell therapy for treating various neurological disorders such as, neurodegenerative diseases, spinal cord injury, and stroke. NES cells’ therapeutic potential lies in their ability to differentiate into multiple cell types and restore neuronal function. However, in order for this therapeutic potential to be exploited several modifications must be made. One of the important aspects for upscaling production is that the cell culture media must adhere to good manufacturing practice (GMP) guidelines. Currently existing media is supplemented with a complex of nutrients called “B27”, which has a composition derived from non-human origin and is chemically undefined. Thus, there is a need to replace B27 with a suitable alternative that promotes NES cell expansion and proliferation. We have formulated a chemically defined media and assessed it by a morphological evaluation of the produced cell cultures, and characterization of the medium candidates using molecular biology techniques such as proliferation and viability assay, and flow cytometry. Overall, this work demonstrates the potential of the optimized defined media for culturing NES cells and lays the groundwork for further research to optimize its formulation and fully understand its impact on NES cell behavior and function. (Less)
- Popular Abstract
- Envision replacing damaged brain cells with healthy ones, treating diseases like strokes, and spinal cord injuries. Scientists are working on this future with special cells called Neuroepithelial Stem Cells (NES). These wonder-cells can transform into different brain cell types, however there is an obstacle – growing them in large numbers. My research focused on developing a special recipe, free of animal-derived ingredients (Causing variability in cells cultures and contaminated with dangerous components) to grow these NES cells efficiently.
NES cells are often grown in a broth-like mixture called media, but a key part of this media comes from animal sources and is poorly defined. This makes it difficult to produce NES cells in large... (More) - Envision replacing damaged brain cells with healthy ones, treating diseases like strokes, and spinal cord injuries. Scientists are working on this future with special cells called Neuroepithelial Stem Cells (NES). These wonder-cells can transform into different brain cell types, however there is an obstacle – growing them in large numbers. My research focused on developing a special recipe, free of animal-derived ingredients (Causing variability in cells cultures and contaminated with dangerous components) to grow these NES cells efficiently.
NES cells are often grown in a broth-like mixture called media, but a key part of this media comes from animal sources and is poorly defined. This makes it difficult to produce NES cells in large quantities and consistently. I investigated a new, defined media recipe to support the growth of NES cells and examine if they would thrive.
The results were exciting! The NES cells seemed to adjust to the new recipe after a while, forming structures like miniature rosettes. Interestingly, the count of cells grown in the new media seemed less, suggesting a possible difference in growth patterns. We also investigated various NES cell stemness (ability to self-renew) and health signs. The findings were mixed, where some markers showed similar activity in both media (optimized media and currently used media), while others suggested slight changes in the NES cells grown on the new media. This highlights the need for further investigation.
Overall, my project provides a steppingstone towards developing a defined media for growing NES cells. This paves the way for future research to optimize the recipe and fully understand its impact on NES cell’s function. With more work, this could lead to compliance with required cell therapy guidelines (Good Manufacturing Practice) and large-scale production of healthy and functional NES cells, a crucial step towards regenerative therapies for various neurological disorders. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9162329
- author
- Musaad, Rana LU
- supervisor
-
- Alice Hägg LU
- organization
- course
- KIMM01 20241
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9162329
- date added to LUP
- 2024-06-12 14:25:42
- date last changed
- 2024-06-12 14:25:42
@misc{9162329,
abstract = {{Neuroepithelial stem (NES) cells derived from induced pluripotent stem cells (iPSCs), hold promise as a novel cell therapy for treating various neurological disorders such as, neurodegenerative diseases, spinal cord injury, and stroke. NES cells’ therapeutic potential lies in their ability to differentiate into multiple cell types and restore neuronal function. However, in order for this therapeutic potential to be exploited several modifications must be made. One of the important aspects for upscaling production is that the cell culture media must adhere to good manufacturing practice (GMP) guidelines. Currently existing media is supplemented with a complex of nutrients called “B27”, which has a composition derived from non-human origin and is chemically undefined. Thus, there is a need to replace B27 with a suitable alternative that promotes NES cell expansion and proliferation. We have formulated a chemically defined media and assessed it by a morphological evaluation of the produced cell cultures, and characterization of the medium candidates using molecular biology techniques such as proliferation and viability assay, and flow cytometry. Overall, this work demonstrates the potential of the optimized defined media for culturing NES cells and lays the groundwork for further research to optimize its formulation and fully understand its impact on NES cell behavior and function.}},
author = {{Musaad, Rana}},
language = {{eng}},
note = {{Student Paper}},
title = {{A Systematic Approach to Media Development for Efficient Expansion of Neuroepithelial Stem Cells in a GMP Setting}},
year = {{2024}},
}