M Soleimani, S Nadri , R Izadpanah ,
Volume 66, Issue 4 (7-2008)
Abstract
Background: MSCs have been isolated from a variety of mammals by the plastic adherence method. However, this method can be problematic due to the unwanted growth of hematopoietic cells and non-MSCs. The potential of MSCs to differentiate along multiple lineages is the key to the identification of stem cell populations in the absence of molecular markers. In the present study, we describe a homogeneous population of MSCs from mouse bone marrow isolated using an improved plastic adherence method that employs frequent medium change (FMC) at the initial hours of harvested bone marrow cell culture.
Methods: Balb/c mice were sacrificed and whole bone marrow cells were aspirated from the femur and tibia and then cultivated in six-well plates. After 3-4 hours of culture, old medium was removed and fresh medium was added. FMC was performed every eight hours over a 72 hour period. When primary cultures became nearly confluent, the first passage was performed. These cells were then used for further examination. To investigate their mesenchymal nature, the cells were allowed to differentiate into mesenchymal lineages and examined at each passage up to the tenth passage for surface antigens by flow cytometry.
Results: We achieved purified populations of fibroblast-like cells in the two weeks after culture initiation. The cells were capable of differentiating into osteocytes and adipocytes. Isolated MSCs were reactive to the CD44, Sca-1, and CD90 cell surface markers. MSCs were negative for hematopoietic surface markers such as CD34, CD11b, CD45, CD31, CD106, CD117 and CD135.
Conclusions: This protocol provides an efficient isolation of homogeneous populations of MSCs from mouse bone marrow.
Seyed Khalil Pestehei, Mahdieh Ghiasi, Seyed-Hassan Emami-Razavi ,
Volume 81, Issue 7 (10-2023)
Abstract
Human mesenchymal stromal cells are multipotent cells capable of differentiating into the mesenchymal lineage that can be isolated from bone marrow and adipose tissue or from umbilical cord blood and fetal tissues. Among the widely characterized in vitro properties, MSCs show strong anti-proliferative and anti-inflammatory effects on immune responses Exosomes derived from mesenchymal stem cells derived from different tissues are promising cell-free treatments for tissue damage repair. Exosomes serve as a potential portal for cell-free drug delivery systems, as these drugs possess the properties of the parent cell from which they are derived. Extracellular vesicles (EVs) play key roles in cell biology and may provide new clinical diagnostics and therapies. Exosomes, called extracellular vesicles (EcVs), are present in almost all cells, tissues, and body fluids. They contribute to intercellular signaling and maintain tissue homeostasis. The biogenesis of exosomes starts in the endosomal system. Researchers have identified 9769 proteins, 2838 miRNAs, 3408 and 1116 lipids present in exosome of mRNA cargo. Isolation of exosomes from cells, tissues and body fluids follows a different pattern. Exosomes interact with receptor cells through their surface receptor molecules and ligands and are internalized into receptor cells through micropinocytosis and phagocytosis. This varies depending on the origin of the EV, its physiological and pathological state, and even the exact site of cellular release. The composition of the protein inside can also indicate the presence of disease pathologies such as cancer or inflammatory diseases; However, exosomes also contain a number of common proteins as well as proteins involved in vesicle formation. Advanced technologies in regenerative medicine have caused researchers to use exosomes isolated from mesenchymal stem cells (MSCs) with high regeneration ability in diseases. Exosome cargo plays a key role in diagnosis and treatment by controlling the disease process. Various studies in laboratory conditions have shown the effectiveness and therapeutic potential of exosomes in cancer, neurodegenerative, cardiovascular and orthopedic diseases. This article describes the therapeutic role and potential of exosomes derived from mesenchymal stem cells, as well as the necessary precautions for their processing.
