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. 2023 Jan 27;33(1):10834.
doi: 10.4081/ejtm.2023.10834.

Better isolation, proliferation and differentiation of human adipose-derived mesenchymal stem cells using human serum

Atena Sadat Ghoreishi  1 , Ehsan Iranmanesh  2 , Hosna Rastegarpouyani  3 , Sogand Mokhtarian  4 , Zahra Poshtchaman  5 , Zeinab Sadat Javadi  6 , Alireza Khoshdel  7
Affiliations

Better isolation, proliferation and differentiation of human adipose-derived mesenchymal stem cells using human serum

Atena Sadat Ghoreishi et al. Eur J Transl Myol. .

Abstract

Mesenchymal stem cells have many applications in medicine. Attention to the proliferation and differentiation of stem cell differentiation is an important issue. The aim of this study was to investigate the possibility of optimal isolation, proliferation, and differentiation of adipose tissue-derived mesenchymal stem cells (ADSCs) using human serum. Human serum (HS) was obtained from the venous blood of eight healthy individuals. The rate of proliferation and differentiation of ADSCs and expression of surface markers was assessed by flow cytometry. Bone differentiation was assessed using Alizarin Red staining. Data were analyzed using statistical software. Over time, HS showed more proliferation than fetal bovine serum (FBS) -enriched cells (p <0.05). Differentiation of ADSCs cells ls in HS-enriched medium is faster and more pronounced than differentiation in the control group. The expression of surface markers in the medium containing HS was the same as the medium containing FBS where the expression levels of CD105 and CD95 were found to be positive and the expression of CD34 and CD45 was negative. Due to the better proliferation of adipose tissue-derived mesenchymal cells in the medium containing HS than FBS, it is suggested that human serum be used in future clinical studies. Also, HS is healthier, safer, more accessible, and more affordable than FBS.

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Conflict of interest statement

I confirm that I have read the Journal’s position on ethical publication issues and affirm that this report is consistent with those guidelines.

So far, many research studies have been conducted inthe field of medicine, most of which are in the field of common and infectious diseases. For instance, various studies have been conducted in the field of cancer, and in recent years, with the outbreak of coronavirus, studies have mainly gone in this direction. However, the treatment of these diseases has always been very significant. Additionally, much research has been conducted regarding medicine and health statistics to prove the importance of the healthcare system. One of these methods is cell therapy using stem cells and extracellular vesicles derived from these cells. Stem cells have many applications in medicine., Mesenchymal stem cells are commercially used for three reasons: first, they can be located in areas of inflammation and tissue damage, such as wounds or tumors. Second, mesenchymal stem cells have immunosuppressive properties and can modulate innate and adaptive immune responses. Third, these cells have the ability to support other cells with paracrine factors. Among these, mesenchymal stem cells are of great interest.

Mesenchymal stem cells have the power to differentiate into different cells and are also able to become undifferentiated cells similar to their own. These cells have the ability to differentiate into connective tissue lineages, including adipocytes, chondrocytes, and osteocytes. Mesenchymal stem cells are capable of regenerating their divisions for a long time and at the same time maintaining their differentiation power., The clinical application of stem cells in the treatment of brain injury, Parkinson's, myocardial infarction, incomplete osteogenesis, fracture healing, and tendon rupture, as well as treatments of joint, liver and other disorders have been well demonstrated.,

One of important sources of potent mesenchymal stem cells is adipose tissue, which is considered a very suitable source of stem cells., Adipose tissue-derived stem cells (ADSCs) are easily harvested and multiplied, are effective and non-invasive, and have a high capacity for multiplication and proliferation. These types of autologous sstem cells significantly reduce donation complications.,

On the other hand, litterature findings suggest that use of animal serum in clinical cases of cell therapy is not a completely safe choice. Indeed, animal serum may cause immune reactions and infections. Providing the safest and most cost-effective serum with optimal properties and greater safety for cell therapy is an important issue. Human serum (HS) seems to be a suitable alternative, because of to its high availability, safety and rapid collection. Human serum contains many useful proteins, including growth factorsand immunoglobulins, inducing a wide variety of positive effects on cell culture.

Studies on ADSC have found that HS can be used to produce and proliferate fat stem cells, reporting that human ADSC proliferation in growth medium containing HS is higher than in culture medium containing fetal bovine serum (FBS). In fact, proteins and other nutrient factors in human serum affect the expression of inflammatory agents in stem cells.

Quantitative studies have been performed on various aspects of the advantage of human serum over animal serum, so the aim of this study was to confirm or not the effects of human serum for better isolation, proliferation and differentiation of ADSC in cell culture medium.

Figures

Fig 1.
Fig 1.
Difference in the number of cells on consecutive days of culture in both mediums enriched with HS and FBS
Fig 2.
Fig 2.
Specific staining of cultured cells containing medium enriched with HS and FBS. A. Control group (FBS), b) The first group of human serum (day 9), c) The second group of human serum (day 12), d) The third group of human serum (15th day), (10x magnification).
Fig 3.
Fig 3.
Specific oil red O staining on cultured cells containing media enriched with HS and FBS. A) Control group (FBS), b) Human serum group.
See this image and copyright information in PMC

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