After years of experience, proficient researchers often have a few stem cell culture tips and tricks up their sleeves. That’s because stem cells come with challenges as unique as their source individuals. For example, is the cell donor older? At any age, does the cell donor have specific disease processes in place? If the answer to either (or both) is “yes”, then this article is for you.
The diseases that most notably affect the quality of stem cells that are harvested from donors include diabetes, arthritis and osteoarthritis, glucocorticoid imbalances, diseases of the cardiovascular system, obesity and hypercholesterolemia, cancer, and aplastic anemia. It is also likely that aged or diseased individuals will have fewer stem cells that can be cultivated. When culturing stem cells from donors, it is important to understand how aging and disease can affect your culture outcomes.
Sometimes, clinicians work with mesodermal cells that have been reprogrammed into mesenchymal stem cells. Mesenchymal cells are support cells found in adults in tissues from bone marrow, adipose tissue, and peripheral blood. Mesenchymal cells can also be found in neonatal cells such as the placenta, plus the umbilical cord and cord blood. Note that StemCultures does not use embryonic stem cells.
Often, when mesenchymal cells are sourced from donors, they come with shades of age or disease. That’s an unfortunate consequence of the fact that the adult human who needs stem cells may be aging, ailing, or both. Typically, it’s more likely that the clinician can successfully address age issues over disease issues.
Research has demonstrated an improvement of mesenchymal stem cells through hypoxic pretreatment, heat shock, or restriction of calories. There are also growth factors and cytokines that can be utilized to better maintain stem cell cultures. Prior research using mice has demonstrated that the combination of aged mice serum with that of younger mice resulted in overall freshened serum (Choudhery et. al, 2021).
In vitro expansion of stem cells is another challenge. This expansion routinely results in lower-quality cell production, but clinicians can overcome this. If not addressed, the expanded culture of stem cells will include those affected by faulty paracrine activity, meaning that the cells do not signal one another properly. In addition, there can be difficulties with differentiation and migration. Stem cell tips and tricks utilizing the same strategies of hypoxia, heat shock, and caloric restriction will help such cultures.
While most clinicians focus on the use of stem cells to treat disease processes, stem cells can also be used to determine the efficacy of new therapies, such as pharmacologic treatment. Cells can be extracted from donor patients, reprogrammed and expanded, then used as a personalized treatment method. Instead of exposing the individual to a new medication that might bring deleterious effects to the patient, the medication can be tested using a line of stem cells. For someone with a neurologic disorder, for example, the clinician could determine if the medication improved the condition without substantial side effects.
When expanding cell lines from donors, another way to overcome limitations with aged or diseased lines is to have the highest quality reagents and additives to promote the best growth. Some data show that these feeder additives can supplement the use of human cell feeder layers, which are subject to contamination from associated pathogens and mycoplasmas. Contaminants can also interfere with the monitoring of the culture, which emphasizes the importance of growing the culture with minimal interruptions for feeding or adjustment.
StemCultures media additives consist of controlled-release growth factor products that will benefit your cell cultures by stabilizing growth factor levels and reducing the number of feedings. Today, they’re used by iPSC core facilities, academic institutions, and biotechnology companies. Choose StemBeads® or DISC Devices™ for stem cell culture media, or contact StemCultures for more information.
Once the donor stem cell lines are ready to be transported, often time scientists will freeze the vial of cells for shipping. Freezing your stem cell culture can result in the differentiation failure or breakdown of the culture into single cells. It’s best to use a reagent of 90 percent bovine serum albumin (BSA) and 10 percent dimethyl sulfoxide (DMSO). StemCultures also recommends a freshly prepared freezing medium, although others have proceeded successfully with a medium that was prepared in advance and stored in a dark environment for a month. (Castro-Viñuelas, Rocio et al, 2021 )
