PAKISTAN VETERINARY JOURNAL

Pak Vet J, 2026, 46(4) 930-939

TBP and HPRT1 are the Most Suitable Reference Genes for Normalization of mRNA Expression in Feline Gonadal Tissue-Derived Mesenchymal Stem Cells

Sang-Yun Lee^1,†,‡, Sanghyeon Park^2,†, Tae-Hyun Park^1,†, Chan-Hee Jo², Seong-Ju Oh², Chae-Yeon Hong², Yong-Ho Choe², Yeon-Woo Jeong³, Hyeon-Jeong Lee², Eun-Yeong Bok⁴, Won-Jae Lee⁵, Sung-Lim Lee² and Young-Bum Son^1,*

^*1Department of Obstetrics, College of Veterinary Medicine, Chonnam National University, 300 Yonbongdong, Buk-gu, Gwangju 61186, Republic of Korea; ²Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju, 52828, Republic of Korea; ³Department of Companion Animal and Animal Resources Science, Joongbu University, Geumsan 32713, Republic of Korea; ⁴Division of Animal Diseases & Health, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Republic of Korea; ⁵Department of Obstetrics, College of Veterinary Medicine, Kyungpook National University, Daegu 41566, Republic of Korea. ‡Current affiliation: Department of Biology Education, Korea National University of Education, Cheongju 28173, Republic of Korea.

*Corresponding author: ybson@jnu.ac.kr

Abstract

Mesenchymal stem cells (MSCs), especially those derived from gonadal tissues that are easy to collect during routine sterilization procedures, provide an ethically viable source of feline MSCs. Considering MSCs are potential candidates for regenerative medicine, reporting therapeutic effects suggests that further research to develop more effective therapy using feline gonadal MSCs is needed for clinical application. Gene expression analysis and therapeutic potential of MSCs can be analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Reliable interpretation of results requires accurate normalization. Hence, this study focused on determining the highest stability of candidate reference genes and accurate gene expression determination in ovary-derived MSCs (O-MSCs) and testis-derived MSCs (T-MSCs). O-MSCs and T-MSCs were retrieved from three female and three male felines, and then their stemness properties, which included spindle-like morphology, specific surface markers, and trilineage differentiation capacity, were characterized. To rank the nine candidate reference gene stability and pairwise variation stability, geNorm and NormFinder were used. In terms of overall analysis, TBP and HPRT1 had the lowest stability values (<0.03), whereas GUSB, RPL7, and ACTB had higher stability values (>0.04). When stable reference genes were utilized for normalization, statistically significant differences in OCT4 expression were observed between O-MSCs and T-MSCs (P<0.05), which were not observed with unstable reference genes. This provides a methodological framework by which future qRT-PCR-based studies can be conducted on feline gonadal MSCs. The potential therapeutic applications of these cells can also be studied.

To Cite This Article: Lee SY, Park S, Park TY, Jo CH, Oh SJ, Hong CY, Choe YH, Jeong YW, Lee HJ, Bok EY, Lee WJ, Lee SL and Son YB, 2026. TBP and HPRT1 are the most suitable reference genes for normalization of mRNA expression in feline gonadal tissue-derived mesenchymal stem cells. Pak Vet J, 46(4): 930-939. http://dx.doi.org/10.29261/pakvetj/2026.070

Abstract

ISSN 0253-8318 (Print)ISSN 2074-7764 (Online)

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)