Laboratory Research Projects
THERAPEUTIC ACTION OF MESENCHYMAL STEM CELLS, PLATELETS + LEUKOCYTES
Overview: This study is intended to characterize the role of each cell type in the treatment of damaged intervertebral disc. In particular, the data collected from this work will be provide valuable evidence and support for future clinical approaches used when treating patients with injured discs.
- Mesenchymal stem cell (MSC) populations are being expanded in vitro (in the lab) for use int his study, a process requiring specific performance factors in the cell be determined to ensure suitability for use.
- Four-color flow cytometry is used to verify the stromal cell population comprises MSCs using a validated protocol.
- Collection, decellurizatiion and disinfection of porcine nucleus pulposus has been achieved.
- Leukocyty-rich and leikocyte-poor platelet preparations are being collected and validated.
- In vitro analysis of cell growth and biochemical changes in the nucleus pulposus are being initiated.
- Methods for tracking and differentiating MSC from platelets ans leukocytes in vitro are underway.
Preliminary Findings: Unique to our data, we are able to describe a mechanism of action by which mesenchymal stem cells respond to nucleous pulposus material in the presence of platelets and leukocytes.
Goal: In-depth data and statistical analysis is currently underway. manuscript preparation has begun, with the intention of submitting our findings for peer-review publication by early 2019.
PROTOCOL DEVELOPMENT FOR QUALITY ASSURANCE
Flurorescent Micropy: It is essential to evaluate every bone marrow aspirate (BMAC) product for quality, which is achieved by determining the number of total nucleated cells concentrated in the product before re-injection.The use of fluorescent viability dyes allow us to quantify the total nucleated cells in a BMAC with an unparalleled level of accuracy and efficiency. This approach has reduced our cell quantification protocol run time from 45 minutes to 10 minutes.
Flow Cytometry: Mesenchymal stem cells are a distinct cell type among the nucleated cells in the bone marrow. it is beneficial to determine that these stem cells of interest exist at the expected frequency in the final product. Using a highly sensitive, four color (label) flow cytometer, we are able to detect the mesenchymal stem cells as individual particulates to ensure their frequency is within a range we believe to be therapeutic.
Stem Cell Painting: A small fraction of the final BMAC product is cultured in our research lab to ensure that the stem cells are of adequate health to self-renew and populate a new environment. This is carried on for several generations and ultimately, purified populations of MSC can be collected and used for laboratory projects.
EVALUATION OF CROSS TALK BETWEEN MESENCHYMAL STEM CELL SUBPOPULATIONS
We have identified unique growth kinetics of MSCs derived from a distinct portion of the bone marrow. We are interested in the potential of combining these MSCs with those traditionally harvested and evaluate the regenerative potential of the mixture.
- Currently, we are collecting data on the different growth kinetics of the two cell types independent of one another.
- Future steps will be to evaluate the cells in co-culture, where one of the cell types is labeled with a fluorescent tracker and is distinguishable from others.
- We suspect there is a soluble form of communication that is inherently robust in one of the stem cell types. We intend to evaluate the potential of these factors to influence the growth kinetics of other cells without the physical cell type being present in the mixture. This will translate to a modified therapeutic approach where we are able to reply on stem cell survival based on a specific mixture of the cellular components.
Reduce Pain. Increase Mobility + Function. Improve Quality of Life.
All the research conducted through the Napa Medical Research Foundation, a 501(c)(3) nonprofit organization, is fully funded through generous donations received from individuals and family foundations.