Background: Research on stem cells has become one of the most promising and advanced scientific topics. Nevertheless, there is a lack of this type of research in the field of forensic diagnostics. Age estimation is one of the main worked-on issues in forensic practice and research. Still, there is a need to explore new complementary reliable methods. This study aimed to detect the possibility of age estimation via age-associated alteration of RIF-1 (Replication timing regulatory factor 1), TCL1A (T-cell leukemia/lymphoma-1A), and TERT (Telomerase reverse transcriptase) gene expression in rats bone marrow stem cells. Methods: Isolation of bone marrow mesenchymal stem cells from rats were undertaken at different ages; 2, 4, 6, 8, 10, and 12 weeks old. RNA extraction was performed then total RNA was reverse transcribed to cDNA to detect the quantitative expression of RIF-1, TLC1A, and TERT by using the qRT-PCR then immunoblotting. Results: There were statistically significant increases in RIF1 and TCL1A mRNA and protein expression and a decrease in TERT mRNA and protein expression with age. Conclusion: Our results showed that the mRNA and protein expression of RIF-1, TCL1A, and TERT in bone marrow cells of rats could be useful indicators for age estimation in forensic practice.