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"Penelitian mengenai sel punca serta aplikasinya di bidang biomedis telah mengalami perkembangan yang pesat. Namun, penggunaan sel punca embrionik dalam penanganan medis menimbulkan kekhawatiran terkait bioetika penggunaan embrio manusia, penolakan sel punca oleh sistem imun tubuh pasien, dan risiko terbentuknya teratoma. Hal tersebut menimbulkan urgensi untuk membuat sel punca pluripoten dari sel somatik yang telah terdiferensiasi. Metode induced pluripotent stem cells dapat dimanfaatkan untuk membuat sel-sel punca dari sel-sel yang telah terdiferensiasi. vesikel ekstraseluler yang dihasilkan oleh sel-sel iPSC juga menjanjikan dalam inovasi aplikasi iPSC minim risiko. Untuk dapat mengaplikasikan vesikel ekstraseluler iPSC sebagai penanganan medis dengan dosis yang tepat, diperlukan uji sterilitas dan karakterisasi profil metabolit vesikel ekstraseluler dari iPSC. Tujuan dari penelitian ini adalah untuk mengetahui sterilitas sampel vesikel ekstraseluler iPSC SCTE FKUI dan mengetahui karakteristik profil metabolit dari sampel vesikel ekstraseluler iPSC yang didapatkan di SCTE FKUI. Metode penelitian yang digunakan mencakup uji sterilitas menggunakan Respiratory Flow Chip Kit dan analisis profil metabolit menggunakan Ultra-Performance Liquid Chromatography-Mass Spectrometry yang dilanjutkan dengan analisis menggunakan perangkat lunak MassLynx 4.1. Hasil menunjukkan bahwa sampel vesikel ekstraseluler iPSC koleksi FKUI steril terhadap 23 patogen infeksi saluran pernapasan akut, dan memiliki profil metabolit berupa 12 metabolit dengan karakteristik beragam.

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The research on stem cells and their applications in the biomedical field has been advancing, but the usage of embryonic cells had provoked controversies regarding the bioethics of extracting human embryonic tissues and poses a risk of rejection from patients’ immune system and the possibility of teratoma formation. Hence, the urgency to utilize a stem cell derived from terminally differentiated cells comes to surface. The iPSC method can be used to reprogram terminally differentiated cells back into cells with pluripotency. Extracellular vesicles secreted by iPSC are also promising for biomedical treatment. In order to use extracellular vesicles for biomedical treatment with a suitable dosage, it is necessary to confirm the sterility and characterize the extracellular vesicles of iPSC used. This research is aimed to test the sterility and analyze the metabolite profile of vesikel ekstraseluler iPSC samples from SCTE FKUI collection. Respiratory Flow Chip Kit is used as sterility test. Ultra-performance liquid chromatography-mass spectrometry is used to to analyze metabolite profile. The analysis of the metabolite profile is conducted with Masslynx 4.1. Result of the sterility test shows that the vesikel ekstraseluler iPSC collection was sterile from 23 pathogens of acute respiratory infections. Moreover, it is confirmed from LCMS analysis that the sample contains 12 metabolites with various characteristics.

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"Bradiaritmia seperti total AV blok dan disfungsi nodus SA (NSA) memerlukan alat pacu jantung permanen (APJP), namun APJP memiliki potensi komplikasi akut dan jangka panjang. Sel punca mesenkimal (SPM) dapat mengatasi kerusakan nodus sinoatrial (NSA) dan nodus atrioventrikel (NAV), namun transplantasi SPM secara langsung pada hewan model memiliki angka keberhasilan rendah. Untuk mengatasi masalah tersebut, diperlukan sel punca yang telah terdiferensiasi menjadi pacemaker-like cell seperti sel NSA atau NVA sehingga siap ditransplantasikan ke area disfungsi atau blokade. Penelitian ini bertujuan untuk mendiferensiasi SPM asal jaringan adiposa (SPMA) menjadi pacemaker-like cells. Penelitian ini menggunakan desain eksperimental dan dilakukan pada bulan Februari 2022 sampai Maret 2023 di IMERI-FKUI. Terdapat 5 kelompok penelitian, yaitu: 1) kultur SPMA tanpa intervensi (kontrol), 2) kultur SPMA yang didiferensiasi menjadi kardiomiosit, 3) kultur SPMA yang didiferensiasi menjadi kardiomiosit dan ditransfeksi gen Tbx3 (TBX), 4) kultur SPMA yang didiferensiasi menjadi kardiomiosit dan diberikan small molecules SB431542, serta 5) kultur SPMA yang didiferensiasi menjadi kardiomiosit, ditransfeksi gen Tbx3 dan diberikan small molecule SB431542 (TBX+SM). Pemeriksaan ekspresi gen penanda pacemaker-like cells (Tbx3, Cx30, Cx40, Cx43, HCN1, HCN3, HCN4, dan KCNN4) menggunakan metode qRT-PCR pada kelompok TBX, SM, dan TBX+SM menunjukkan peningkatan ekspresi gen Tbx3, Cx30, HCN1, HCN3, HCN4 dan KCNN4 yang berbeda bermakna terhadap kelompok kardiomiosit (p ≤ 0,001) sedangkan penurunan ekspresi gen Cx40 dan Cx43 berbeda bermakna dibandingkan kelompok kardiomiosit (p < 0,001). Eskpresi protein Tbx3 dan Cx30 menggunakan ELISA pada kelompok TBX, SM, dan TBX+SM berbeda bermakna terhadap kelompok kardiomiosit (p ≤ 0,001). Gambaran ekspresi protein Tbx3 dan Cx30 menggunakan metode imunofluoresensi menunjukkan pendaran positif pada kelompok TBX, SM, dan TBX+SM. Morfologi elektrofisiologis dengan patch clamp menunjukkan gambaran potensial aksi khas pacemaker-like cells pada kelompok TBX, SM, dan TBX+SM dinilai dari rasio action potential duration90/action potential duration50 (APD90/APD50). Disimpulkan transfeksi gen Tbx3, pemberian small molecules SB431542, dan kombinasi keduanya mampu mendiferensiasikan SPMA menjadi pacemaker-like cells.

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Bradyarrhythmias, such as total AV block and SA node dysfunction, require a permanent pacemaker (PPM); however, PPM has the potential for acute and long-term complications. Mesenchymal stem cells (MSC’s) can repair damaged sinoatrial (SAN) and atrioventricular (AVN) nodes; however, transplantation of MSCs into animal models has a low success rate. To overcome this problem, it is necessary to have stem cells that differentiate into pacemaker-like cells, such as SAN or AVN cells, so that they can be transplanted to areas of dysfunction or blockage. This study aimed to differentiate MSC’s from adipose tissue (AMSC) into pacemaker-like cells. This study used an experimental design and was conducted from February 2023 to March 2023 at the IMERI-FKUI. There were five study groups, namely:1) AMSC cultures without intervention (control), 2) AMSC cultures that differentiated into cardiomyocytes, 3) AMSC cultures that differentiated into cardiomyocytes and transfected with the Tbx3 (TBX) gene, 4) AMSC cultures that differentiated into cardiomyocytes and administered SB431542, and 5) AMSC culture, which differentiated into cardiomyocytes, were transfected with the Tbx3 gene and administered SB431542 (TBX+SM). RT-qPCR expression of pacemaker-like cell marker genes (Tbx3, Cx30, Cx40, Cx43, HCN1, HCN3, HCN4, and KCNN4) in the TBX, SM, and TBX+SM groups showed increased expression of Tbx3, Cx30, and HCN1., HCN3, HCN4, and KCNN4, which differed significantly in the cardiomyocyte group (p ≤ 0.001), whereas the decrease in Cx40 and Cx43 gene expression was significantly different compared to that in the cardiomyocyte group (p < 0.001). ELISA of Tbx3 and Cx30 protein expression in the TBX, SM, and TBX+SM groups was significantly different from that in the cardiomyocyte group (p ≤ 0.001). Immunofluorescence analysis of Tbx3 and Cx30 protein expression showed a positive correlation in the TBX, SM, and TBX+SM groups. The electrophysiological morphology with the patch clamp showed a typical action potential picture of pacemaker-like cells in the TBX, SM, and TBX+SM groups, as assessed by the ratio of action potential duration90/action potential duration50 (APD90/APD50). It was concluded that transfection of the Tbx3 gene, administration of the small molecule SB431542, and a combination of both could differentiate SPMA into pacemaker-like cells."

"This volume looks at induced pluripotent stem (iPS) cells, mature cells that have been genetically reprogrammed so that they return to their embryonic state."

"Latar belakang: Hati kelinci yang dideselularisasi sebagai perancah untuk kultur organoid hati telah menunjukkan hasil yang menjanjikan dalam meningkatkan viabilitas dan fungsinya. Penelitian ini bertujuan untuk mengembangkan organoid dari kokultur sel yang dapat mendukung fungsi hati. Pemanfaaatan perancah hati yang dideselularisasi untuk mempertahankan viabilitas dan fungsionalitas hepatosit dan mengevaluasi organoid hati manusia yang ditransplantasikan ke model hewan coba dan mengetahui respons yang dimediasi sel imun. Metode: Sel hepatosit yang berasal dari iPSC manusia dikokultur dengan tiga sel lain untuk membentuk organoid hati. Delapan belas belas kelinci putih berusia 3 bulan digunakan dalam percobaan ini dan dibagi menjadi empat kelompok: Kelompok sham-operated (n = 3), kelompok ligasi duktus biliaris (n = 6), kelompok eksperimen dengan ligasi saluran empedu diikuti oleh transplantasi organoid hati (kelompok jangka pendek, (n=5); kelompok jangka panjang, (n=4).Hasil: Pada penelitian ini dilakukan analisis survival menggunakan metode Kaplan-Meier (KM) untuk menentukan probabilitas kumulatif kelangsungan hidup dari kejadian kematian pada kedua kelompok dengan dan tanpa transplantasi organoid hati. Hasil tes log-rank menunjukkan bahwa kemungkinan bertahan hidup secara keseluruhan antara kedua kelompok yang menerima perlakuan berbeda. (p=0,003). Kelompok jangka pendek menunjukkan peningkatan fungsi hati seperti albumin, CYP3A, dan tingkat AST yang lebih rendah daripada kelompok jangka panjang. Hati kelompok jangka pendek menunjukkan tingkat deposisi kolagen yang lebih rendah.Kesimpulan: Transplantasi organoid hati kokultur manusia dalam perancah hati yang dideselularisasi ke hewan yang diligasi duktus biliaris dapat mendukung kelangsungan hidup hewan dan fungsi hati untuk jangka pendek. Studi ini menyoroti potensi transplantasi organoid hati untuk mendukung fungsi hati jangka pendek. Namun, fungsi dan penolakan organoid hati dapat membatasi penggunaan pada jangka panjang.

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Background: Decellularized native liver scaffolds as a platform for liver organoid culture have shown promising results in improving their viability and function. This research aims to develop cocultured liver organoids that can recapitulate liver functions, utilize a decellularized native liver scaffold to maintain the viability and functionality of hepatocytes and evaluate human liver organoids transplanted into animal models to support liver function in two periods categories and immune-mediated response.

Methods: The hepatocyte-like cells derived from the human iPSCs were cocultured with three other cells to form liver organoids. Eighteen 3-month-old New Zealand White Rabbits were used in the experiment, divided into four groups: A sham-operated group (n=3), a bile duct ligation group (n=6), an experimental group with biliary duct ligation followed by liver organoid transplantation (short-term group, n=5; long-term group, n=4).

Results: We performed a survival analysis using the Kaplan-Meier (KM) method to determine the cumulative probability of survival from death events in both groups with and without liver organoid transplantation. The log-rank test results indicated a notable variation in the overall likelihood of survival between the two groups receiving different treatments. (p=0.003). The short-term group exhibited improved liver functions such as albumin, CYP3A, and lower levels of AST than the long-term group. The livers of the short-term group showed lower levels of collagen deposition.

Conclusions: Transplanting human coculture liver organoids in decellularized native liver scaffold into bile duct ligated animals could support the animal's survival and hepatic function for the short term. This study highlights the potential of liver organoid transplantation for short-term liver support. However, the functionality and rejection of liver organoids may limit their long-term use."

"Sel punca mesenkim (MSC) dan sel punca pluripoten terinduksi (iPSC) telah dilaporkan mampu berdiferensiasi menjadi hepatosit secara in vitro dengan berbagai tingkat maturasi hepatosit. Sebuah metode sederhana untuk proses deselulerisasi perancah hati telah dikembangkan oleh Fakultas Kedokteran Universitas Indonesia. Penelitian ini bertujuan untuk mengevaluasi diferensiasi hepatosit dari iPSC dibandingkan dengan MSC dalam perancah hati yang dideselularisasi. Langkah pada penelitian ini adalah mengkultur iPSC dan MSC, mendeselularisasi hati kelinci, menyemai kultur sel ke dalam perancah, dan mendiferensiasikan menjadi hepatosit selama 21 hari dengan protokol Blackford yang dimodifikasi. Pemeriksaan dilakukan dengan pewarnaan Haematoxylin Eosin (HE), Masson Trichrome (MT), imunohistokimia (IHK) albumin dan cytochrome 3A4 (CYP3A4). Ekspresi gen albumin, cytochrome P450 (CYP450), dan cytokeratin-19 (CK-19) dianalisis menggunakan qRT-PCR. Pemeriksaan scanning electron microscope (SEM) dan immunofluorescence (IF) marker hepatocyte nuclear factor 4 alpha (HNF4-α) dan CCAAT/enhancer-binding protein alpha (CEBPA) dilakukan. Diferensiasi hepatosit dari iPSC dalam perancah hati yang dideselulerisasi dibandingkan dengan diferensiasi hepatosit dari MSC dalam perancah hati yang dideselulerisasi menunjukkan pembentukan sel tunggal dan kapasitas adhesi pada perancah yang lebih sedikit, dan penurunan tren ekspresi albumin dan CYP450 yang lebih rendah. Jumlah penyemaian sel awal yang lebih rendah menyebabkan hanya beberapa iPSC menempel pada bagian-bagian tertentu dari perancah hati yang dideselularisasi. Injeksi jarum suntik manual untuk reselulerisasi yang tidak merata menciptakan pola pembentukan sel tunggal oleh hepatosit dari diferensiasi iPSC di perancah hati yang dideselulerisasi. Hepatosit dari diferensiasi MSC memiliki kapasitas adhesi lebih tinggi ke perancah hati yang dideselulerisasi yang mengarah pada peningkatan tren ekspresi albumin dan CYP450. Penurunan ekspresi gen CK-19 lebih banyak terjadi pada diferensiasi hepatosit dari iPSC. Hasil tersebut dikonfirmasi oleh adanya sinyal positif protein HNF4-α dan CEBPA dengan pemeriksaan IF yang menunjukkan hepatosit yang dewasa. Kesimpulan dari penelitian ini adalah diferensiasi hepatosit dari iPSC pada perancah hati yang dideselularisasi lebih dewasa dengan adhesi sel-matriks ekstraseluler lebih rendah, distribusi sel spasial saling berjauhan, dan ekspresi albumin dan CYP450 lebih rendah dibandingkan dengan diferensiasi hepatosit dari MSC pada perancah hati yang dideselularisasi.

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Mesenchymal stem cells (MSC) and induced pluripotent stem cells (iPSC) have been reported able to differentiate to hepatocyte in vitro with varying degree of hepatocyte maturation. A simple method to decellularized liver scaffold has been established by Faculty of medicine Universitas Indonesia.

This study aims to evaluate hepatocyte differentiation from iPSCs compared to MSCs in decellularized liver scaffold. iPSCs and MSCs were cultured, rabbit liver were decellularized, cell cultures were seeded into the scaffold, and differentiated into hepatocytes for 21 days with modified Blackford protocol. Haematoxylin-Eosin (HE), Masson Trichrome (MT), immunohistochemistry (IHC) albumin and CYP3A4 was performed. Expression of albumin, cytochrome P450 (CYP450) and cytokeratin-19 (CK-19) genes were analyzed using qRT-PCR. Scanning electron microscope (SEM) and immunofluorescence (IF) examination of hepatocyte nuclear factor 4 alpha (HNF4-α) and CCAAT/enhancer-binding protein alpha (CEBPA) marker was performed.

Hepatocyte differentiated iPSCs compared with hepatocyte differentiated MSCs in decellularized liver scaffold single–cell–formation and lower adhesion capacity in scaffold, and decrease trends of albumin and CYP450 expression. Lower initial seeding cell number causes only a few iPSCs to attach to certain parts of decellularized liver scaffold. Manual syringe injection for recellularization abruptly and unevenly create pattern of single–cell–formation by hepatocyte differentiated iPSCs in the decellularized liver scaffold. Hepatocyte differentiated MSCs have higher adhesion capacity to decellularized liver scaffold that lead to increase trends of albumin and CYP450 expression. CK-19 expression gene diminished more prominent in hepatocyte differentiated iPSCs.

These results were confirmed by the presence of HNF4-α and CEBPA positive signal protein with IF examination, showing mature hepatocyte.The conclusion of this study is hepatocyte differentiated iPSCs in decellularized liver scaffold differentiation is more mature with lower cell-extracelullar matrix adhesion, spatial cell distribution far from each other, and lower albumin and CYP450 expression than hepatocyte differentiated MSCs in decellularized liver scaffold."

"Stem Cell Labeling for Delivery and Tracking Using Noninvasive Imaging provides a comprehensive overview of cell therapy imaging, ranging from the basic biology of cell therapeutic choices to the preclinical and clinical applications of cell therapy. It emphasizes the use of medical imaging for therapeutic delivery/​targeting, cell tracking, and determining therapeutic efficacy. The book first presents background information and insight on the major classes of stem and progenitor cells. It then describes the main imaging modalities and state-of-the-art techniques that are currently employed for stem cell tracking. In the final chapters, leading scholars offer clinical perspectives on existing and potential uses of stem cells as well as the impact of image-guided delivery and tracking in major organ systems. Through clear descriptions and color images, this volume illustrates how noninvasive imaging is used to track stem cells as they repair damaged tissue in the body. With contributions from some of the most prominent preclinical and clinical researchers in the field, the book helps readers to understand the evolving concepts of stem cell labeling and tracking as the field continues to move forward."

"This book discusses the various methods to reprogram cells, the control and determination of cell identity, the epigenetic models that have emerged and the application of iPS cell therapy for brain diseases, in particular Parkinson’s disease and Vanishing White Matter (VWM).​"

"Human pluripotent stem cells, including human embryonic stem cells and induced pluripotent stem cells, are a key focus of current biomedical research. The emergence of state of the art culturing techniques is promoting the realization of the full potential of pluripotent stem cells in basic and translational research and in cell-based therapies. This comprehensive and authoritative atlas summarizes more than a decade of experience accumulated by a leading research team in this field. Hands-on step-by-step guidance for the derivation and culturing of human pluripotent stem cells in defined conditions (animal product-free, serum-free, feeder-free) and in non-adhesion suspension culture are provided, as well as methods for examining pluripotency (embryoid body and teratoma formation) and karyotype stability. "

"Terapi penyakit degeneratif menggunakan sel punca mesenkim (SPM) dikembangkan dengan pendekatan seluler ataupun dengan conditioned medium (CM) yang mengandung faktor pertumbuhan dan vesikel ekstraseluer (VE). Sel punca kanker merupakan populasi kecil sel dalam jaringan kanker yang berkaitan dengan resistensi terapi. Belum diketahui dampak VE SPM tali pusat terhadap kepuncaan sel kanker payudara. Penelitian ini bertujuan menganalisis dampak pemberian VE SPM tali pusat terhadap kepuncaan sel kanker payudara. VE diisolasi dengan kromatografi kolom; diidentifikasi dengan mikroskop konfokal dan transmission electron microscope. Internalisasi VE oleh sel kanker payudara dikonfirmasi dengan mikroskop konfokal. Analisis viabilitas sel pasca kokultur VE dilakukan menggunakan trypan blue exclusion assay, ekspresi mRNA OCT4 dengan qRT-PCR, ekspresi protein OCT4 dengan Western Blot, aktivitas enzim ALDH dengan ALDEFLUOR™. Hasil, VE SPM tali pusat berhasil diisolasi serta diidentifikasi. Derajat internaliasasi VE oleh ketiga jenis sel kanker payudara berbeda. VE 5% meningkatkan viabilitas ketiga jenis sel serta ekspresi mRNA OCT4 sel MCF7 dan ALDH+. Tingkat ekspresi protein OCT4 sel MCF7 dan ALDH+ berbanding terbalik dengan peningkatan konsentrasi VE. VE 5% meningkatkan ekspresi protein OCT4 sel MDA-MB-231. VE 5% meningkatkan aktivitas ALDH ketiga sel kanker payudara. Pada VE 10%, aktivitas ALDH sel MDA-MB-231 dan MCF7 menurun, namun pada sel ALDH+ meningkat. Kesimpulan, pemberian VE SPM tali pusat dengan konsentrasi yang berbeda memberikan dampak berbeda terhadap kepuncaan berbagai sel kanker payudara, berkaitan dengan regulasi ekspresi OCT4 dan aktivitas ALDH.

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Therapy of degenerative diseases using umbilical cord mesenchymal stem cells (UCMSCs) are currently developed either using the cell or the conditioned medium containing extracellular vesicles (EVs). Cancer stem cells are a minor subpopulation of cells within cancerous tissue that had been associated with therapy resistance. This study aimed to investigate the effect of EVs secreted by UCMSC (UCMSC-EVs) on the stemness of human breast cancer cells. UCMSC-EVs were isolated using SEC, then identified using confocal microscope and TEM. UCMSC-EV uptake by MDA-MB-231, MCF7, and ALDH+ cells was analyzed by confocal microscope. The viability of co-cultured breast cancer cells was determined using trypan blue exclusion assay, mRNA and protein expression of OCT4 as well as ALDH activity were analyzed qRT-PCR, Western Blot, and ALDEFLUOR™, respectively. As the result, UCMSC-EVs were successfully isolated and identified. The internalization ability of each type of breast cancer cell seemed different. Notably, 5% EVs increased the viability of those three cells. Five percent of EVs increased the mRNA expression of OCT4. On MCF7 and ALDH+ cells, the higher the EVs concentration given, the lower expression of OCT4 protein was. Supplementation of EVs 5% increased the ALDH activity of cells. In conclusion, supplementation of UCMSC-EVs in different concentrations gives different impacts in terms of stemness that was correlated with OCT4 and ALDH regulation within the treated cells."

"Pemahaman karakteristik sel punca kanker merupakan salah satu cara untuk menemukan terapi yang tepat untuk mengobati penyakit kanker. Penelitian ini bertujuan untuk menemukan pengaruh lingkungan mikro yang dihasilkan oleh sel fibroblas normal dan kanker terhadap pluripotensi sel punca kanker payudara. Sel fibroblas dan sel punca kanker payudara masing-masing dikultur dengan menggunakan medium kultur DMEM high glucose. Kemudian sel punca kanker diko-kultur dengan sel fibroblas, baik sel fibroblas normal maupun kanker. Pengukuran pluripotensi dilakukan dengan 2 cara, yaitu pengukuran ekspresi penanda permukaan CD44+/CD24+ dengan spektrofluorometer dan pengukuran ekspresi SOX2 dengan menggunakan reverse transcription-polymerase chain reaction. Hasil pengukuran menunjukkan bahwa pluripotensi sel punca kanker payudara menurun pada sel punca kanker yang diko-kultur dengan sel fibroblas, baik fibroblas normal maupun kanker, namun, ekspresi penanda permukaan dan SOX2 pada sel punca kanker yang diko-kultur dengan sel fibroblas kanker lebih tinggi dibandingkan dengan yang diko-kultur dengan sel fibroblas normal. Dari hasil ini, kami menyimpulkan bahwa lingkungan mikro yang dihasilkan sel fibroblas normal dan kanker mampu menurunkan tingkat pluripotensi sel punca kanker payudara sehingga lingkungan mikro dapat dimanfaatkan sebagai sarana untuk menghilangkan sel punca kanker.

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Understanding and figuring out the characteristics of cancer stem cells is believed as a way to find a perfect therapy in treating cancer disease. This research aims to find out the effect of the microenvironment provided by either normal fibroblast cells or cancer fibroblast cells toward the pluripotent characteristics of breast cancer stem cells. Both the fibroblast cells and the cancer stem cells were cultured independently using DMEM high glucose. The cancer stem cells were then cocultured into the fibroblast cells, both normal and cancer cells. The pluripotent characteristics were measured using two methods; expression of CD44+/CD24+ cell surface markers using fluorescent spectroscopy and expression of SOX2 using reverse transcription - polymerase chain reaction. Results showed that the expression of both CD44+/CD24+ cell surface markers and SOX2 decreased in breast cancer stem cells co-cultured with the fibroblast cells, whereas the expression in the cancer stem cells co-cultured with cancer fibroblast cells were higher than those co-cultured with the normal fibroblast cells. From the results, we suggest that the microenvironment created by either normal fibroblast cells or cancer fibroblast cells could decrease the pluripotent characteristics of breast cancer stem cells, hence microenvironment can be used as a tool to eradicate cancer stem cells."