pf-mop

Oral Presentations

M.S. Student Oral Presentations

MOP01

Thanatotranscriptome Analysis of Prostate Tissue after Death

Mariah Tolbert1, Sheree J. Finley2, Jessica Carter1, and Gulnaz T. Javan1,*

1Forensic Science Program, Department of Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104; 2Department o Physical/Forensic Sciences, Alabama State University, Montgomery, Alabama 36104. *Email:gjavan@alasu.edu

Prostate glands are among the last internal organs to decay during human decomposition; however, this phenomenon is still an enigma. Thanatotranscriptome (thanatos-, Greek for death) is the study of postmortem mRNA transcript abundance profiles in human tissues. It has been found that mRNA is stable in specific human tissues up to 96 hrs (Bauer et al., 2003; Partemi et al., 2010). Previous findings from the Javan Thanatos Lab have shown that RNA is a suitable and stable molecule in postmortem liver samples up to 48 hours (Javan et al., 2015). We hypothesized that there are also significant changes in mRNA transcript abundances in prostate tissues from Italian homicide and suicide victims. The goal of the current study is to identify apoptotic biomarkers that show measurable gene expression profiles regarding time of death. Prostate tissue was collected from five cadavers at various times of death along with a control with a PMI of 24 hrs. RNA was extraction and cDNA was synthesized and quantified. Results of total RNA extraction revealed concentrations of greater than 50 ng/_l. The cDNA was profiled using apoptosis-related PCR Array. PCR Array results showed that at 38 hrs after death, a majority of genes for apoptosis induction and positive regulation were over-expressed more than at 60 hrs. In conclusion, our study demonstrated that up-regulation of apoptotic genes in prostate tissues occurs and is measurable after death. Our findings reveal active postmortem gene expression profiles which may play critical roles in time of death determinations and organ transplantation.

Bauer M, Gramlich I, Polzin S, and Patzelt D (2003) Quantification of mRNA degradation as possible indicator of postmortem interval—a pilot study. Legal medicine, 5, 220-227.

Partemi S, Berne PM, Batlle M, Berruezo A, Mont L, Riuró H, et al. (2010) Analysis of mRNA from human heart tissue and putative applications in forensic molecular pathology. Forensic science international, 203, 99-105.

Javan GT, Can I, Finley SJ, and Soni S (2015) The apoptotic thanatotranscriptome associated with the liver of cadavers. Forensic science, medicine, and pathology, 11, 509-516.

MOP02

Enhanced Hepatogenic Differentiation of Bone Marrow Derived Mesenchymal Stem Cells on Liver ECM Hydrogel

Breaunna Primeaux1, Bo Wang1,*, Wuwei Li2, Derrick Dean1, Manoj K. Mishra3, and Kennedy S. Wekesa3

1Biomedical Engineering Program, Alabama State University, Montgomery, Alabama 36014; 2Department of Oral and Maxillofacial Surgery, School of Stomatology, Dalian Medical University, Liaoning 116044, P.R. China; 3College of Science, Mathematics and Technology, Alabama State University, Montgomery, Alabama 36104. *E-mail: bwang@alasu.edu

BM-MSC is a promising alternative cell source to primary hepatocytes because of their ability to differentiate into hepatocyte-like cells. However, their inability to differentiate efficiently and potential to turn into myofibroblasts restrict their applications. This study developed a plate coating from the liver extracellular matrix (ECM) and investigated its ability in facilitating the BM-MSCs proliferation, hepatic differentiation, and hepatocyte-specific functions during in vitro culture. After 28-day culture, BM-MSCs on the ECM coating showed hepatocyte-like morphology, and certain cells took up LDL. Synthesis of albumin, urea, and AFP, as well as expression of certain hepatic markers, in cells cultured on ECM were higher than cells cultured on non-coated and Matrigel-coated plates. mRNA levels of CYP3A4, albumin, CK18, and CYP7A1 in cells on ECM coating were significantly higher than cells cultured on the non-coating environment. In conclusion, viability and hepatogenic differentiation of BM-MSCs cultured on both Matrigel and ECM coating were significantly enhanced compared to those cultured on non-coated plates. Moreover, the liver ECM coating induced additional metabolic functions relative to the Matrigel coating. The liver ECM hydrogel preserves the natural composition, promotes simple gelling, induces efficient stem cell hepatogenic differentiation, and may have uses as an injectable intermedium for hepatocytes.