Impact of Adipose Tissue Depot Harvesting Site on the Multilineage Induction Capacity of Male Rat Adipose-Derived Mesenchymal Stem Cells: An In Vitro Study

Hussein M El-Husseiny; Masahiro Kaneda; Eman A Mady; Tadashi Yoshida; Ahmed S Doghish; Ryou Tanaka

doi:10.3390/ijms24087513

Impact of Adipose Tissue Depot Harvesting Site on the Multilineage Induction Capacity of Male Rat Adipose-Derived Mesenchymal Stem Cells: An In Vitro Study

Int J Mol Sci. 2023 Apr 19;24(8):7513. doi: 10.3390/ijms24087513.

Authors

Hussein M El-Husseiny^{1

2}, Masahiro Kaneda³, Eman A Mady⁴, Tadashi Yoshida⁵, Ahmed S Doghish^{6

7}, Ryou Tanaka¹

Affiliations

¹ Laboratory of Veterinary Surgery, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai Cho, Fuchu-shi 183-8509, Tokyo, Japan.
² Department of Surgery, Anesthesiology, and Radiology, Faculty of Veterinary Medicine, Benha University, Moshtohor, Toukh 13736, Elqaliobiya, Egypt.
³ Laboratory of Veterinary Anatomy, Division of Animal Life Sciences, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai Cho, Fuchu-shi 183-8509, Tokyo, Japan.
⁴ Department of Animal Hygiene, Behavior, and Management, Faculty of Veterinary Medicine, Benha University, Moshtohor, Toukh 13736, Elqaliobiya, Egypt.
⁵ Department of Applied Biological Science, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
⁶ Department of Biochemistry, Faculty of Pharmacy, Badr University in Cairo (BUC), Badr City 11829, Cairo, Egypt.
⁷ Department of Biochemistry, Faculty of Pharmacy (Boys), Al-Azhar University, Nasr City 11651, Cairo, Egypt.

Abstract

Recently, substantial attention has been paid toward adipose-derived mesenchymal stem cells (AdMSCs) as a potential therapy in tissue engineering and regenerative medicine applications. Rat AdMSCs (r-AdMSCs) are frequently utilized. However, the influence of the adipose depot site on the multilineage differentiation potential of the r-AdMSCs is still ambiguous. Hence, the main objective of this study was to explore the influence of the adipose tissue harvesting location on the ability of r-AdMSCs to express the stem-cell-related markers and pluripotency genes, as well as their differentiation capacity, for the first time. Herein, we have isolated r-AdMSCs from the inguinal, epididymal, peri-renal, and back subcutaneous fats. Cells were compared in terms of their phenotype, immunophenotype, and expression of pluripotency genes using RT-PCR. Additionally, we investigated their potential for multilineage (adipogenic, osteogenic, and chondrogenic) induction using special stains confirmed by the expression of the related genes using RT-qPCR. All cells could positively express stem cell marker CD 90 and CD 105 with no significant in-between differences. However, they did not express the hematopoietic markers as CD 34 and CD 45. All cells could be induced successfully. However, epididymal and inguinal cells presented the highest capacity for adipogenic and osteogenic differentiation (21.36-fold and 11.63-fold for OPN, 29.69-fold and 26.68-fold for BMP2, and 37.67-fold and 22.35-fold for BSP, respectively, in epididymal and inguinal cells (p < 0.0001)). On the contrary, the subcutaneous cells exhibited a superior potential for chondrogenesis over the other sites (8.9-fold for CHM1 and 5.93-fold for ACAN, (p < 0.0001)). In conclusion, the adipose tissue harvesting site could influence the differentiation capacity of the isolated AdMSCs. To enhance the results of their employment in various regenerative cell-based therapies, it is thus vital to take the collection site selection into consideration.

Keywords: AdMSCs; adipose stem cells; differentiation; fat depots; harvesting site; male rats.

MeSH terms

Adipose Tissue / metabolism
Animals
Cell Differentiation
Cells, Cultured
Male
Mesenchymal Stem Cells* / metabolism
Osteogenesis*
Rats
Subcutaneous Fat

Grants and funding

This work was supported by a full scholarship from the Egypt-Japan Education Partnership call 4 (EJEP-4) from the Ministry of Higher Education of the Arab Republic of Egypt. Moreover, it was supported by the WISE Program: Doctoral Program for World-Leading Innovative and Smart Education of Tokyo University of Agriculture and Technology (TUAT) granted by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.