|Rizal Rizal||Biomedical Engineering, Department of Electrical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16424, Indonesia|
|Rahimi Syaidah||Department of Histology, Faculty of Medicine, Universitas Indonesia, Depok, West Java, 16424, Indonesia|
|Evelyn Evelyn||Undergraduate Program in Biomedical Engineering, Department of Electrical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16424, Indonesia|
|Alif Hafizh||Undergraduate Program in Biomedical Engineering, Department of Electrical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16424, Indonesia|
|Josh Frederich||Undergraduate Program in Biomedical Engineering, Department of Electrical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, West Java, 16424, Indonesia|
Wharton’s jelly mesenchymal stem cells (WJ-MSCs) is one of the best sources of mesenchymal stem cells (MSCs) that suggest both embryonic and adult stem cell characteristics. Before being applied in clinical application, the isolated MSCs should be tested to assess their quality, including differentiation capacity, phenotype characterization, and morphological appearance. This research aims to quantify the differentiation capacity of WJ-MSCs isolated using explant method. The WJ-MSCs cells were grown out from Wharton’s jelly tissue and the isolated cells adhered in T25 plastic flask. The isolated cells expressed high amount of MSC surface marker which are CD105 (99.97±0.06%), CD73 (99.97±0.06%), and CD90 (99.12±0.25%). The cells can be differentiated into adipocytes, chondrocytes, and osteocytes. The quantification showed that the amount of mineralization in osteoblastogenesis, production of lipid droplet in adipogenic differentiation, and production of glycosaminoglycan in chondrogenesis were noticeably higher in differentiated cells than non-differentiated cells. In conclusion, the isolated cells fulfill the minimum criteria of MSCs that can be used in research or clinical application. The great differentiation capacity of the cells into osteocytes and chondrocytes indicate that the cells are suitable in bone tissue engineering application, both for research and clinical application.
Adipocytes; Chondrocytes; Differentiation capacity; Osteocytes; WJ-MSCs
Present materials, which are incorporated with osteoinductive properties, are continually developed for bone tissue engineering utilization to generate osteogenesis at the implant site. Graphene, which is generally a monoatomic two-dimensional sheet-like material with sp2-hybridized carbon atoms arranged in a hexagonal or honeycomb-like structure, and its thickness identical to an atom diameter, is one example of these materials with documented pro-osteogenic effects (Hermenean et al., 2016; Kusrini et al., 2019). However, it possesses a challenge to produce the materials (Supriadi et al., 2017). Another example is mesoporous silica nanoparticles (MSN). Osteogenic agents are added to the MSNs augment the bone regeneration process (Narayan et al., 2018). Porous materials are widely used as adsorbents, catalysts, and catalyst support due to their large surface area and pore volume characteristics (Wilson and Mahmud, 2015). However, to increase bone healing recovery, those materials should be combined with multipotent cells that have the properties of high self-proliferation and differentiation into bone-related cells.
Multipotent cells with high capacity of self-proliferation that can be derived from almost all parts of the body, including neonatal byproducts, bone marrow, adipose tissue, and dental tissue, are called Mesenchymal stem cells (MSCs) (Hass et al., 2011; Shivakumar et al., 2019). The MSCs hold a promising potential application for regenerative disease and immunomodulation (Abdallah and Kassem, 2008). These have been approved for the treatment of various diseases such as Crohn-related enterocutaneous fistular disease and graft versus host disease (Galipeau and Sensébé, 2018). The MSCs have also been explored to address several immunological disease (Ghannam et al., 2010), bone and cartilage defects (Krampera et al., 2006), neurological degeneration (Karussis et al., 2010), and cardiovascular diseases (Ranganath et al., 2012).
Several findings suggested that birth byproducts have better proliferation and differentiation capacity (Anzalone et al., 2010; Hass et al., 2011). The MSCs can be isolated from various birth byproducts including amniotic membrane and fluid (Wolbank et al., 2007; Utama, 2018), umbilical cord (Van Pham et al., 2016), Wharton’s jelly tissue (Widowati et al., 2019), and umbilical cord blood (Bieback and Netsch, 2016). Isolated MSCs from neonatal-derived tissues also have both embryonic and adult stem cell characteristics (Arutyunyan et al., 2016).
Wharton’s jelly tissues are part of the umbilical cord that are considered as one of the finest sources of MSCs. The advantages of using these tissues are ethical consideration, their availability, and non-invasive isolation procedure (Hass et al., 2011). Before their clinical application, there are several quality controls to examine the quality of WJ-MSCs. The minimal criteria that have been accepted in both industrial and basic research application has been published by the International Society for Cellular Therapy (ISCT) (Dominici et al., 2006). There are three minimal criteria for MSCs: adherence to plastic, positive (>95%) for CD105, CD73, and CD90, and can be differentiated into osteocytes, chondrocytes, and adipocytes.
The aspects that may affect the differentiation capacity of MSCs are tissue origin, isolation method, culture condition, and cells passage (Ahern et al., 2011; Hass et al., 2011; Nepali et al., 2018; Rizal et al., 2019). They also can be trans-differentiated into ectodermal lineage and endodermal lineage cells, including ?-pancreas (Ullah et al., 2019), neuronal cells (Cortés-Medina et al., 2019), and cardiomyocytes (Arslan et al., 2018). This differentiation capacity makes stem cells prospective for transplantation, thus having the ability to repair many organ disfunctions. In addition, MSCs are able to migrate and differentiate in the area of injury using the ability called homing capacity (Lin et al., 2017; Ullah et al., 2020). These benefits make the research of exploring the potential of MSCs very popular (Zakrzewski et al., 2019).
Differentiation capacity into osteocytes is one of the strengths of WJ-MSCs that can be applied in bone tissue engineering (Ansari et al., 2018). The WJ-MSCs reveal all characteristics of functional osteocytes/osteoblasts due to its osteogenic gene expression, the ability to adhere in scaffold, and expression of extracellular matrix mineralization (Todeschi et al., 2015). They have been successfully transplanted into patients to treat osteonecrosis and exhibited improvement in the joint function and also relieved the pain (Cai et al., 2014).
Compared with bone marrow mesenchymal stem cells (BM-MSCs), the application of WJ-MSCs in bone tissue engineering has several advantages. The isolation procedure of WJ-MSCs are non-invasive because it comes from byproduct waste pain (Wang et al., 2016). The WJ-MSCs also have low immunogenicity that enable us to use these cells in both autologous and allogenic transplantation. When transplanted into human body, WJ-MSCs are protected against lysis by NK cells because these cells express low quantity of primary major histocompatibility class I (MHCI) and class II (MHCII) proteins (Kalaszczynska and Ferdyn, 2015). There are no teratoma formation after transplantation of WJ-MSCs in mice, as well as the patients (Ding, 2015).
Present study strives to quantify the differentiation capacity of MSCs into three different mesodermal cells lineage: adipocytes, chondrocytes, and osteocytes. Because of the heterogeneity of stem cells, the quantification of the quality of stem cells become an important criterion in the quality check of MSCs before being transplanted into human body, and in future all the minimal criteria of stem cells should be measurable to ease the quality control check of MSCs.
The current study indicates that the WJ-MSCs, isolated through explant methods, generate high-quality stem cells that are in line with mesenchymal stem cell criteria. The isolated WJ-MSCs can be differentiated into adipocytes, chondrocytes, and adipocytes. This capacity can be quantified, producing better determination on the quality of stem cells and their role in bone tissue engineering.
This study was supported by a grant from Universitas Indonesia, PUTI Prosiding 2020, contract no. NKB-912/UN.RST/HKP.05.00/2020.
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