Effect of passaging on the stemness of infrapatellar fat pad‑derived stem cells and potential role of nucleostemin as a prognostic marker of impaired stemness

Radhakrishnan,Subathra; Trentz,Omana  Anna; Martin,Catherine  Ann; Reddy,Mettu  Srinivas; Rela,Mohamed; Chinnarasu,Marimuthu; Kalkura,Narayana; Sellathamby,Shanmugaapriya

doi:10.3892/mmr.2019.10268

Effect of passaging on the stemness of infrapatellar fat pad‑derived stem cells and potential role of nucleostemin as a prognostic marker of impaired stemness

Authors:
- Subathra Radhakrishnan
- Omana Anna Trentz
- Catherine Ann Martin
- Mettu Srinivas Reddy
- Mohamed Rela
- Marimuthu Chinnarasu
- Narayana Kalkura
- Shanmugaapriya Sellathamby
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Affiliations: National Foundation for Liver Research, Cell Laboratory, Gleneagles Global Health City, Chennai 600100, India, MIOT Institute of Research, MIOT International, Chennai 600089, India, Institute of Liver Disease and Transplantation, Gleneagles Global Health City, Chennai 600100, India, Crystal Growth Centre, Anna University, Chennai 600025, India, Department of Biomedical Science, Bharathidasan University, Tiruchirappalli 620024, India
Published online on: May 22, 2019 https://doi.org/10.3892/mmr.2019.10268
Pages: 813-829
Copyright: © Radhakrishnan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Infrapatellar fat pad‑derived stem cells (IFPSCs) are emerging as an alternative to adipose tissue‑derived stem cells (ADSCs) from other sources. They are a reliable source of autologous stem cells obtained from medical waste that are suitable for use in cell‑based therapy, tissue engineering and regenerative medicine. Such clinical applications require a vast number of high‑quality IFPSCs. Unlike embryonic stem cells (ESCs), ADSCs and IFPSCs have limited population doubling capacity; however, in vitro expansion of primary IFPSCs through multiple passages (referred to as P) is a crucial step to acquire the desired population of cells. The present study investigated the effect of multiple passages on the stemness of IFPSCs during expansion and the possibility of predicting the loss of stemness using certain markers. IFPSCs were isolated from infrapatellar fat pad tissue resected during knee arthroplasty performed on aged patients (>65 years old). These cells from the stromal vascular fraction were serially passaged to at least to P7, and their stemness characteristics were examined at each passage. It was observed that IFPSCs maintained their spindle‑shaped morphology, self‑renewability and homogeneity at P2‑4. Furthermore, immunostaining revealed that these cells expressed mesenchymal stem cell (CD166, CD90 and CD105) and ESC markers [Sox2, Nanog, Oct4 and nucleostemin (NS)], whereas the hematopoietic stem cell marker CD45 was absent. These cells were also able to differentiate into the three germ layer cell types, thus confirming their ability to generate clinical grade cells. The findings indicated that prolonged culture of IFPSCs (P>6) led to the loss of the stem cell proliferative marker NS, with an increased population doubling time and progression toward neuronal differentiation, acquiring a neurogenic phenotype. Additionally, IFPSCs demonstrated an inherent ability to secrete neurotrophic factors and express receptors for these factors, which is the cause of neuronal differentiation at later passages. Therefore, these findings validated NS as a prognostic indicator for impaired stemness and identified IFPSCs as a promising source for cell‑based therapy, particularly for neurodegenerative diseases.

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