Potential of Conditioned Medium of hATMSCs in Aging Cells Model

Skin aging is caused by the exposure cumulative of ultraviolet radiation, it leads reactive oxygen species (ROS) production in the skin. The conditioned medium of human Adipose Tissue-derived Mesenchymal Stem Cells (hATMSCs) can scavenge free radicals and increase the survival rate of skin cells under oxidative stress. This study examined the protective effects of Conditioned Medium (CM) of hATMSCs in H2O2-induced human skin fibroblast cell line (BJ). The aging cells model using H2O2-induced BJ cells were added CM-hATMSCs in concentrations (0, 10, 30%) and incubated in various time, furthermore BJ cells induced by various H2O2 concentrations (0, 50, 100, 200 µM) incubated for 1 h. The anti-aging potential were measured including viability, ROS and collagen levels in BJ cells which treated CM-hATMSCs. The median inhibitory concentration (IC50) of H2O2 on BJ cells for 1 h incubation was 107.87 μM and 91.25 μM for 10 min incubation. CM-hATMSCs increased the viability on aging model cells. CM-hATMSCs concentration 30% increased the viability of H2O2 50, 100, 200 µM-induced BJ cells. CM-hATMSCs concentration 25% decreased ROS, increased collagen level in H2O2 50, 100, 200 µM-induced BJ cells. CM-hATMSCs increase the viability cells, collagen level and decrease ROS level in aging model cells

OPTIMASI ISOLASI SEL PUNCA DARI JARINGAN LEMAK MANUSIA DAN IMMUNOPHENOTYPING

In the past few decades, attention and research in the field of stem cell are progressing very rapidly. Hospitals in Indonesia have been using stem cells as an alternative to cure some illnesses like diabetes, heart disease, fractures and joints, dental implants, and asthma. Currently, adult stem cells can be obtained not only from the spinal cord and peripheral vessels, but also from fat tissues of the human body, where it can be isolated as adherent stem cells (mesenchymal stem cells). Human adipose tissue is a great source of mesenchymal stem cells. Adipose-derived mesenchymal stem cells (AD-MSCs) are easily isolated, can differentiate into multi-lineage cells and have various clinical application. Consideration of fat tissue as the source of mesenchymal stem cells (MSCs) for autologous tissue engineering is because they are readily available in abundant quantities through minimal invasive procedures, as well as easily cultured and propagated. Besides, the fat tissue is a type of adult stem cells (ASCs) that capable of metamorphosis / transdifferentiation into various strains due to the nature of high plasticity. Therefore it is possible to proliferate and differentiate into the desired direction of the network. Therefore, the aim of this study is to support the government in providing a stem cell production technology that can be used for alternative treatment for diseases that are difficult to cure with chemical treatments such as cancer and diabetes

Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells to Osteoblast Cell in Early, Middle and Late Passages

Human adipose tissue is a great source of mesenchymal stem cells. Adipose-derived mesenchymal stem cells (AD-MSCs) are easily isolated, able to differentiate into multi-lineage cells and have various clinical application. This promising potential will be more valuable for the application of tissue engineering if they can be sequentially subcultured without loss of phenotype and multilineage differentiation ability. Thus, in this study we investigated the ability of human AD-MSCs to differentiate into osteogenic lineage by performing histological staining and studied the osteogenic potential of human AD-MSCs on early (P2, P5), middle (P10) and late passage (P15). Cells were cultured in osteogenic medium for 2 weeks and characterized at 1, 4, 7, 10 and 14 days. During sequential subcultivation, AD- MSCs were able to maintain their osteogenic potential through late passage. AD-MSCs in middle passage exhibited a better osteogenic potential than the early and late passages, while the AD-MSCs from P2 are less differentiated than middle and late passages. The result that was found in this research should be accounted for when developing stem cell-based therapies for clinical application Keywords: Adipose-derived mesenchymal stem cell; Mesenchymal stem cell, Differentiation; Osteoblast cel

POTENSI SEL PUNCA MESENKIM ASAL JARINGAN ADIPOSA MENJADI SEL OSTEOBLAS

Human adipose tissue is a great source of mesenchymal stem cells. Adipose-derived mesenchymal stem cells (AD-MSCs) are easily isolated, can differentiate into multi-lineage cells and have various clinical application. Commonly, the osteogenic differentiation of MSC were induced using culture medium that was supplemented with L-ascorbic acid 2- phosphate (Asc), dexamethasone (Dex) and beta-glycerophosphate (β-GP). The supplement composition of osteogenic medium that was frequently used (OM1 and OM2) was originally generated for bone marrow-derived mesenchymal stem cells, thus the supplement concentration may not be optimal for the differentiation of AD-MSCs. Therefore, in this study we investigated the ability of human AD-MSCs to differentiate into osteogenic lineage using four different osteogenic medium (OM1, OM2, OM3 and OM4). Cells were cultured in osteogenic medium and characterized on day 5. The result suggest that differentiation was enhanced on OM3 and OM4, which have high Asc and low Dex concentrations. Hence higher concentration of Asc, lower concentration of Dex and higher serum concentrations should be used for the osteogenic differentiation of human AD-MSCs in vitro

Optimization of Alginate-Based Encapsulation Utilization For Viability and Stability of The Mesenchymal Stem Cell

In the past few decades, attention and research in the field of stem cell are progressing very rapidly. Hospitals in Indonesia have been using stem cells as an alternative to cure some illnesses like diabetes, heart disease, fractures and joints, dental implants, etc. Currently, adult stem cells can be obtained not only from the spinal cord and peripheral vessels, but also from fat tissues of the human body, where it can be isolated as adherent stem cells (mesenchymal stem cells). Consideration of fat tissue as the source of mesenchymal stem cells (MSCs) for autologous tissue engineering is because they are readily available in abundant quantities through minimal invasive procedures, as well as easily cultured and propagated. It is possible to proliferate and differentiate into the desired direction of the network. Stem cell growth requires conditions to grow such as requiring optimum growing conditions such as an environmental temperature of 37°C and a concentration of 5% CO2. Maintenance of MSCs also requires a subculture process, i.e. the process of moving MSCs from a full culture medium to new media; continuous subculture process can cause changes in MSCs. The viability of stem cells may be disrupted by micro-conditions in wounds such as hypoxia, oxidative stress, and inflammation. Therefore, the purpose of this research was to investigate whether alginate-based encapsulation can increase and maintenance stem cell growth at different temperature by using some concentration of alginate and CaCl2 as the formula. Results shown that alginat with low concentration and CaCl2 100mM is suitable for MSCs growth (as in MTT result shown) at 25°C temperature. This can be due to the MSCs encapsulated can adapt and grow within the alginate microcapsule with low concentration. In addition, the media may also easier to get into the microcapsule alginate

AKTIVITAS SITOTOKSIK EKSTRAK Clerodendrum sp. DAN EKSTRAK Phyllanthus sp. TERHADAP SEL KANKER PARU-PARU (A549) DAN SEL KANKER PARU-PARU YANG DIINFEKSI DENGAN SIMIAN RETRO VIRUS (A549-SRV2) Cytotoxic activity of Clerodendrum sp. and Phyllanthus sp. extract

ABSTRACTCytotoxic activity of Clerodendrum sp. and Phyllanthus sp. from the National Park in Indonesia has been carriedout using MTT assay. In vitro cytotoxicity test of Clerodendrum sp. and Phyllanthus sp. extracts against lung cancercells (A549) showed over 50% cell viability at concentrations of 50 ppm. Cell viability in normal CHO cells was90% at the same concentration. Cytotoxicity of Clerodendrum sp. extract on cancer cells infected with simian retrovirus(A54-SRV) using reverse transcriptase Real Time PCR showed the value of cycle threshold (CT) was 17,83on day1 and 17,72 on day 3. Extract of Phyllanthus sp. showed CT values of 17,32 on day 1 and 15,73 on day 3.Clerodendrum sp. and Phyllanthus sp. showed no growth inhibitory activity of normal CHO cells and lung cancercell A549. Clerodendrum sp.and Phyllanthus sp. extract did not show significant differences in antivirus testingon day 1 between extracts, positive control, and negative control. Phyllanthus sp. extract treatment on day 3 wassignificantly different compared to positive and negative control. Clerodendrum sp. extracts treatment showed nodifference between CT values on day 3 and day 1. Extracts of Phyllanthus sp. showed no potential anticancer andantiviral against lung cancer cells (A549). Clerodendrum sp. extract has potential anticancer activity in cancercells infected with simian retrovirus (A549-SRV-2). ABSTRAKAktivitas sitotoksik ekstrak Clerodendrum sp. dan Phyllanthus sp.yang berasal dari Taman Nasional di Indonesiatelah diuji dengan metode MTT. Pengujian sitotoksisitas in vitro kedua ekstrak terhadap sel kanker paru-paru(A549) menunjukkan viabilitas sel diatas 50% pada konsentrasi 50 ppm. Sedangkan pada sel normal CHO,menunjukkan viabilitas sel sebesar 90% pada konsentrasi yang sama. Aktivitas sitotoksik ekstrak Clerodendrumsp. pada sel kanker yang diinfeksi simian retrovirus (A549-SRV) dengan menggunakan metode reversetranscriptase RT-PCR menunjukkan nilai cycle threshold (CT) sebesar 17,83 pada hari ke-1 dan 17,72 padahari ke-3. Perlakuan menggunakan ekstrak Phyllanthus sp. menunjukkan nilai CT sebesar 17,32 pada hari ke-1dan 15,73 pada hari ke-3. Clerodendrum sp. dan Phyllanthus sp. tidak menunjukkan aktivitas penghambatanpertumbuhan sel normal CHO dan sel kanker paru-paru A549. Pengujian antivirus menunjukkan bahwa padaperlakuan hari ke-1 tidak menunjukkan perbedaan yang nyata antara ekstrak Clerodendrum sp., ekstrakPhyllanthus sp., kontrol positif (lamivudin), dan kontrol negatif. Pada perlakuan hari ke-3, nilai CT dari ekstrakPhyllanthus sp. lebih rendah dan berbeda nyata dibandingkan kontrol positif dan kontrol negatif. Perlakuandengan ekstrak Clerodendrum sp. menunjukkan tidak adanya perbedaan perlakuan pada hari ke-1 dengan harike-3. Ekstrak Phyllanthus sp. tidak berpotensi sebagai antikanker maupun antivirus terhadap sel kanker paruparuA549, sedangkan ekstrak Clerodendrum sp. tidak berpotensi sebagai antikanker pada sel kanker paruparuA54 tetapi berpotensi sebagai antivirus pada sel kanker yang sudah diinfeksi dengan simian retrovirus(A549-SRV-2)

The Encapsulation Effect on Viability of Mesenchymal Stem Cells: Pengaruh Enkapsulasi Terhadap Viabilitas Sel Punca Mesenkimal

Stem cells are potentially used as a regenerative therapy agent. Cell encapsulation is one of the developed methods to utilize Mesenchymal Stem Cells (MSCs) for therapy. This research aimed to study the effect of encapsulation using alginate-CaCl2 towards the viability of hAdMSCs during in vitro culture. Encapsulation of hAdMSCs with alginate-CaCl2 was done using the extrusion method. The viability of hAdMSCs was analyzed with Live/Dead Assay and MTT assay. The results indicated that cultured hAdMSCs within alginate remain alive for 7 days culture period. However, the viability was lower than the reference culture. The absorbances from MTT assay of encapsulated MSCs were lower than the conventional hAdMSCs culture. This result indicated lower metabolic activity of hAdMSCs when cultured in alginate beads

Cytotoxicity Effect of Zingiber officinale, Curcuma aeruginosa and Curcuma xanthorhiza Extracts on Adipose-Derived Stem Cells (ADSCs)

In the past few decades, attention and research in the field of stem cell are progressing very rapidly. Hospitals in Indonesia have been using stem cells as an alternative to cure some illnesses like diabetes, heart disease, fractures and joints, dental implants, and asthma. The purpose of this research was to characterized and to produced stem cells from human subcutaneous adipose tissue (ADSC), which can be used for assessing cytotoxicity effects potency of Zingiber officinale (red ginger), Curcuma aeruginosa (temu ireng) and Curcuma xanthorhiza (temulawak) on ADSC proliferation. The cytotoxic effect was done using MTT assay. The results showed Z. officinale var. and C. xanthorhiza Roxb. extracts have no cytotoxic effect to ADSC at 25 ppm, 50 ppm and 100 ppm concentrations, however the proliferation inhibition of Z. officinale extracts gradually rises to 0.05 % at 200 ppm and slightly fall at 600 ppm, after that it sharply raises unto 74.84% at 1000 ppm. The proliferation inhibition of C. xanthorhiza extracts at 200 ppm is about 28% and significantly increase to 90 % at 1000 ppm which is the highest percentage on the figure. The C. aeruginosa extracts has no inhibition effect to ADSC from 25 ppm until 800 ppm concentration of extract, however it modestly goes up at 1000 ppm reaching 9% of inhibition. Further studies are required to investigate the metabolites in these extracts that are highly correlated with its toxic effects

Growth factors profile in conditioned medium human adipose tissue-derived mesenchymal stem cells (CM-hATMSCs)

Summary: Background: Stem cell-derived by conditioned medium has potential as regenerative agent of skin cells because it contains many growth factors, cytokines and other regenerative biomolecules. Conditioned medium (CM) from human adipose tissue-derived MSCs (CM-hATMSCs) also has been known enriched growth factors that play an important role in epithelial wound repair, reduce wrinkles, and enhance wound healing. Objective: This study was performed to evaluate growth factors in CM such as Transforming Growth Factor (TGF)-β1, TGF-β2, Vascular Endothelial Growth Factor (VEGF)-2, Vascular Cell Adhesion Molecule (VCAM) 1, Epidermal Growth Factor (EGF), Fibroblast Growth Factor (FGF) in CM-hATMSCs treated with Fresh Frozen Plasma (FFP) and non-FFP at various passages. Methods: This study used ELISA method to measure the growth factors in CM-hATMSCs. Results: FGF was the highest modulator among FFP and non-FFP-treated cells at many passages compared to the others growth factor. Both FFP and non-FFP-treated cells showed significant difference (P < 0.05) in some growth factors. However, there was no significant differences in TGF-β1 at passage 3,7,11 and 15 in FFP-treated cells and non-FFP-treated cells. Conclusion: In summary, the highest concentration level found in CM-hATMSCs is FGF both in FFP and non-FFP-treated cells. FGF as growth factor composition in CM-hATMSCs has potential as wound healing and regeneration of cell and it can be used in anti-aging products. Keywords: Conditioned medium, Human adipose tissue derived-mesenchymal stem cells, Growth factor, Fresh frozen plasma, Wound healin