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These paramagnetic immunobeads rapidly isolate high quantities of adipose-derived stem cells while preserving the biological characteristics of the cells that are important for research and clinical use. Stem cells have the unique potential to differentiate along multiple lineages and form various tissue types. Thus, these cells can be utilized for cell-based therapies and tissue regeneration purposes. Diseases ranging from neurodegeneration to heart disease, wound healing to blindness, and many others might one day benefit from stem cell-based therapies. The global market for stem cells was estimated at $9.6 billion in 2019, with an expected compound annual growth rate of 8.2% through 2027.
Typically, stem cells are collected from either bone marrow or adipose tissue. Adipose-derived stem cells offer distinct advantages over their bone marrow-derived counterparts, in that their collection is safer for the patient and less technically challenging. In addition, adipose-derived stem cells contain higher quantities of mesenchymal stem cells to isolate. Mesenchymal stem cells can be differentiated to produce fat, bone, muscle, cartilage, and nerve tissues. However, isolation of stem cells is dependent upon (1) instrumentation that requires electricity and (2) enzymatic solutions. Thus, it is nearly impossible to deliver stem cell-based therapies to the more than 1 billion people worldwide who do not have access to electricity. It is also difficult to obtain FDA approval for stem cell-based therapies that employ enzymatic dissociation, as this process may alter the biological characteristics of the stem cells and impact upon safety and efficacy.
Researchers at the University of Florida have isolated adipose-derived stem cells using paramagnetic immunobeads rather than electricity or enzymatic solutions in a way that preserves full biological characteristics of the cells. Paramagnetic immunobeads, conjugated to selectively bind markers of adipose-derived stem cells, are incubated with lipoaspirate, or fat surgically removed from a patient through common liposuction. By this simple magnetic force, high quantities of these cells can be isolated rapidly. These cells maintain full biological characteristics and are available for research and clinical purposes.
Kit and method for the isolation of adipose-derived stem cells
Immunobeads are conjugated to a panel of antibodies that recognize cell surface markers of adipose-derived stem cells. Incubation of these paramagnetic immunobeads in lipoaspirate allows the beads to bind the cells of interest. Then, a magnetic force pulls the beads and bound cells to the bottom of the incubator, allowing for removal of remaining aspirate and subsequent isolation of these adipose-derived stem cells. In culture, these cells exhibit normal morphology, multipotency, and sphere-forming potential. Stem cells isolated via this technique can be applied as fresh, primary therapeutics, or in autologous assays of toxicology, pharmacology, and disease modeling.
This cell-based microarray allows testing of a wide range of biological or pharmaceutical agents on small numbers of cells from rare populations. Patient-derived cells such as cancer cells, stem cells and precancerous cells are difficult and expensive to obtain. Inefficient existing technologies only allow testing a few agents at a time on small populations of these rare cells, preventing large-scale testing. For example, colon cancer stem cells, recently identified as a potential cause of colon cancer, are of interest to many doctors and researchers who want to test the cells’ response to drugs. Because they are rare, researchers and drug companies can’t conduct large-scale testing on these cells. University of Florida researchers have developed a tool that allows the testing of multiple combinations of biological or pharmaceutical agents on small numbers of cells. Researchers will be able to test more agents without wasting these precious and expensive cells, reducing costs and increasing efficiency. The microarray separates the cells on a glass slide, allowing the user to test many agents at once: each agent targets just a portion of the cells on the slide.
A microarray for the application of multiple agents on a small amount of reagent cells
A microarray is a support (in this case, a glass slide) onto which molecules are attached in a regular pattern for use in biochemical or genetic analysis. In the microarray developed by UF researchers, thin films of timed-release polymer are loaded with testing agents and microarrayed onto solid substrates, providing a background that resists cell adhesion. This tool permits the testing of multiple biological and/or pharmaceutical agents and their various combinations on rare cell populations that are present in small quantities. On a given glass slide more than 1,000 spots can be arrayed, allowing for as many agents to be applied to the cell population of interest. Then, non-adherent or non-reactive cells are removed, leaving separated areas of the cells that showed adherence. Researchers can measure cellular response including proliferation and differentiation, through immunostaining or by using contrast agents.
These cell-based arrays use a small sampling of cells to help isolate and identify a patient’s sensitivity to chemotherapeutic drugs in vitro, potentially personalizing treatment of cancer. The cancer stem cell hypothesis postulates that a minute fraction of cells within a tumor, termed cancer stem cells, possess the tumor-initiating capacity that propels tumor growth. Targeting tumor-initiating cells could be a viable clinical strategy to combating cancer, but these cells are extremely rare, necessitating new methods for rapid and robust screening. Researchers at the University of Florida have developed a miniaturized platform to investigate chemosensitivity of patient-derived tumor-initiating cells using limited cell numbers, potentially personalizing treatment of cancer. These cell-based arrays can predict the success of various combination treatments from one cancer patient to the next using just a small number of cells, thus enabling clinical adoption of personalized approaches to cancer.
Arrays to screen drug efficacy on rare cell populations, such as patient-derived colon cancer stem cells
Cancer stem cells with the greatest tumor-initiating and metastatic potential are exceptionally rare and difficult to isolate. The various embodiments of the arrays in this platform can perform chemosensitivity screens on patient-derived colon cancer stem cells using just a limited number of cells. University of Florida researchers found the results of using these structures and arrays indicate that colon cancer stem cells vary considerably in their response to drugs; this chemosensitivity screening on patient-derived cells can lead to valuable information regarding chemotherapy decisions. In addition to facilitating personalized medicine for treating colon cancer, any cancer stem cell or rare cellular population in which identifying responsiveness to drug combinations is paramount can adopt the same approach.
