Embryonic stem cells (ESCs) hold immense potential for treating a wide range of diseases and injuries. These pluripotent cells, derived from the inner cell mass of blastocysts, possess the remarkable ability to differentiate into any cell type in the human body.
The isolation and maintenance of ESCs require meticulous laboratory techniques and specialized materials to ensure their viability and potency. Here, Creative Biolabs delves into the essential steps, materials, and considerations involved in the isolation and maintenance of ESCs.
To establish an optimal microenvironment for embryonic stem cell growth and maintenance, prepare the culture medium by combining DMEM supplemented with FBS, LIF, and penicillin-streptomycin solution. Ensure sterility by filtering the medium.
Obtain blastocysts from fertilized mouse embryos and transfer the blastocysts to a petri dish containing sterile PBS. Carefully remove the zona pellucida surrounding the blastocysts. Transfer the zona-free blastocysts into a new petri dish containing trypsin solution and incubate.
Stop the trypsin activity by adding an equal volume of culture medium. Centrifuge the cell suspension at low speed and resuspend the cell pellet in fresh culture medium. Plate the embryonic stem cells on tissue culture dishes coated with an appropriate extracellular matrix component.
Place the culture dishes in a cell culture incubator. Regularly replenish the culture medium every 24-48 hours to maintain optimal cell growth. Monitor the embryonic stem cells for morphological changes and evaluate their pluripotency through various assays. To prevent spontaneous differentiation, supplement the culture medium with LIF. Passage the embryonic stem cells by treating them with trypsin and splitting them into new culture vessels as they reach confluency.
You can get stem cell isolation services by contacting us directly.
Reference
For Research Use Only. Not For Clinical Use.
