Joshua Hauser and Dr. Sandra Burnett, Microbiology and Molecular Biology
Successfully culturing and identifying bone marrow cells is a critical skill when doing stem cell, or hematopoietic research. This skill was especially important in Dr. Sandra Burnett’s research lab, with whom I conducted this research. Since the proper culture and identification of the cells was a vital part of any results or conclusions that were made in the lab, we deemed it imperative to determine whether or not we would be able to develop training protocols that would result in better and more consistent results. I strongly believed that this project would immediately impact our current and future research using the MAFIA model (transgenic mice exhibiting macrophage depletion) and our bone marrow transplant research by allowing us to simplify the data collection process and establish a training regimen for future student and faculty researchers. This will aid BYU and other researchers and educators across the country who study hematopoieses using similar methods.
After performing a literature search I realized that there were many articles that attempted to describe the process of using the methyl-cellulose medium to culture bone marrow cells, but these articles were written to a general science audience and the procedures did not detail the exact steps taken to achieve the stated results. I also found that there were no articles concerning the aspect of the research that we were aiming to study. No one had reported any sort of formal training materials or guidelines. The companies that produce and supply the methyl-cellulose medium provide instructions, but these are simply documents to be downloaded and read through. It was not a training course in the least.
In the spring of 2007, Dr. Burnett and I approached members of the Biology 241 class, that I was teaching under her supervision, to determine whether or not we could obtain enough volunteers to perform the project. These students, since they had no prior experience with our techniques, were to be our study group. Our control group was comprised of members of our current research lab who may or may not have had extensive experience with methyl-celluslose culturing, but most likely had performed one more tasks that were related to it. We emailed questionairres to those that were interested and inquired about academic history, time commitment, major, anticipated post-graduate course, and research experience. After reviewing the forms we recieved from the students, Dr. Burnett and I evaluated the applications and selected 16 students to be included in the study group, and 16 to be included in the control group. Students in the control group would not be given any additional training, while the study group was given 8, 1-hour lectures, free access to myself and Dr. Burnett, and an instructional DVD.
During the Winter Semester of 2007 I began to develop a DVD that covered each step of the process in detail. The DVD covered sterile technique, proper equipment, safety procedures, dissection, tissue preparation, culturing, culture evaluation, culture plucking, cytospin, and microscopic evaluation of the cells. Dr. Burnett and I also began to prepare a lecture schedule and lecture materials.
In May of 2007, Dr. Burnett and I began to present the lectures to the study group. We covered the same topics described by the DVD, but took the time to answer questions and to go over the complicated steps more than once in order to ensure that the study group would be as well prepared as possible. The study group was invited to attend and observe other experiments in the lab and were encouraged to ask question and familiarize themselves with the equipment and locations of the materials in the lab.
Following completion of the training course, all members of the research study were given a euthanized mouse and expected to perform the entire culturing and identification procedures from start to finish. Participants were given forms and were to fill them out as they went, taking care to note the amount of time each step took, record the results, and answer questions in order to determine if the students understood why certain steps were performed.
I hypothesized that the students who had participated in the training group would perform significantly better than the students who had received no training. On a whole, the participants from the study group were able to finish the tasks successfully, took less time than the other group to do so, made fewer mistakes, and were better able to answer conceptual questions about the process. Roughly two-thirds of the control group did not obtain usable results, while only one student from the study group failed to produce viable, uncontaminated cultures. The raw data is still in the process of being gleaned for more advanced statistical analysis. After this takes place Dr. Burnett and I plan to publish this research and perhaps present it at a conference of our future choosing.
The research project was not without its stumbling blocks. It was very difficult keep track of all the participants and all of the paperwork. It would have been beneficial to streamline the number of forms used and used a better tracking system. It was also difficult stage procedures between scheduled lab work, classes, and work schedules. I probably should have left myself a bigger window for the practical aspect of the project.
I am very grateful to the ORCA office for funding this project. It was at times stressful and tedious, but it allowed me to make great strides in my personal and academic development and gave me invaluable experience as I work on my Masters in Public Health and for my future goals of obtaining an MD/PhD combined degree.