Derivation, characterization, and in vitro cell regeneration of canine white adipose tissue-derived mesenchymal stem cells obtained from a mesenteric region

The Journal of Basic and Applied Zoology

Dew Biswas¹  Ajeet Kumar Jha²  Shyamal Kanti Guha³  Anirban Mandal⁴ 

[1]Department of Life Science, Presidency University, 700073, Kolkata, India

[2]Department of Veterinary Gynaecology and Obstetrics, West Bengal University of Animal and Fishery Sciences, 700037, Kolkata, India

[3]Stem Cell Laboratory, Department of Veterinary Clinical Complex, West Bengal University of Animal and Fishery Sciences, 700037, Kolkata, India

[4]Stem Cell Laboratory, Department of Veterinary Clinical Complex, West Bengal University of Animal and Fishery Sciences, 700037, Kolkata, India

[5]Department of Microbiology, Mrinalini Datta Mahavidyapith, Birati, 700051, Kolkata, India

关键词: Canine mesenteric white adipose tissue-derived mesenchymal stem cells;    Stromal vascular fraction;    Cell regeneration;    Wound healing assay;    Trilineage differentiation;   

DOI  :  10.1186/s41936-021-00222-1

来源: Springer

【 摘 要 】

BackgroundThe study was conducted to assess the characterization, differentiation, and in vitro cell regeneration potential of canine mesenteric white adipose tissue-derived mesenchymal stem cells (AD-MSCs). The tissue was harvested through surgical incision and digested with collagenase to obtain a stromal vascular fraction. Mesenchymal stem cells isolated from the stromal vascular fraction were characterized through flow cytometry and reverse transcription-polymerase chain reaction. Assessment of cell viability, in vitro cell regeneration, and cell senescence were carried out through MTT assay, wound healing assay, and β-galactosidase assay, respectively. To ascertain the trilineage differentiation potential, MSCs were stained with alizarin red for osteocytes, alcian blue for chondrocytes, and oil o red for adipocytes. In addition, differentiated cells were characterized through a reverse transcription-polymerase chain reaction.ResultsWe observed the elongated, spindle-shaped, and fibroblast-like appearance of cells after 72 h of initial culture. Flow cytometry results showed positive expression for CD44, CD90, and negative expression for CD45 surface markers. Population doubling time was found 18–24 h for up to the fourth passage and 30±0.5 h for the fifth passage. A wound-healing assay was used to determine cell migration rate which was found 136.9 ± 4.7 μm/h. We observed long-term in vitro cell proliferation resulted in MSC senescence. Furthermore, we also found that the isolated cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages.ConclusionsMesenteric white adipose tissue was found to be a potential source for isolation, characterization, and differentiation of MSCs. This study might be helpful for resolving the problems regarding the paucity of information concerning the basic biology of stem cells. The large-scale use of AD-MSCs might be a remedial measure in regenerative medicine.

【 授权许可】

CC BY