Isolation of Wharton's Jelly Mesenchymal Stem Cells and their Derived Exosomes/Small Extracellular Vesicles

Abstract

In the field of regenerative medicine, mesenchymal stem cells (MSCs) have sparked particular attention. MSCs from the umbilical cord (UC) tissue, referred to as Wharton's jelly, have been particularly well-established in a variety of applications. In comparison to other MSC sources, Wharton's jelly MSCs (WJ-MSCs) offer a number of benefits and are isolated from UC, which is typically disposed of as medical waste. WJ-MSCs, therefore, do not raise any ethical issues similar to those pertaining to embryonic stem cells. A simple and reproducible explant technique was used to isolate WJ-MSCs from UC tissue. Within five to ten days of isolation, the WJ-MSCs emerge as adherent cells with a fibroblastic shape. After being isolated, the cells reach about 80% confluency in 14-18 days. Following that, in passage 3, the isolated WJ-MSCs was fully characterized by inducing them to differentiate along the adipogenic, osteogenic, and chondrogenic mesenchymal lineages. Additionally, immunophenotyping for negative cluster of differentiation (CD) surface markers, including CD34, CD14, and HLA-DR, as well as distinctive MSC surface markers like CD105, CD90, CD73, and CD44 were tested. Furthermore, precipitating reagent was used in a straightforward approach to isolate exosomes/small extracellular vesicles (sEVs) from the conditioned medium of those WJ-MSCs. After being isolated, exosomes/sEVs were observed under a transmission electron microscope, and particle size analysis was used to calculate their average size. In summary, WJ-MSCs provide a readily available, non-invasive, renewable supply of stem cells that may be used as factories to produce exosomes/sEVs. These WJ-MSCs and their derived exosomes/sEVs can be used in a wide array of therapeutic and downstream research applications.