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Aplikasi Metode Whole Chromosome Painting untuk Mendeteksi Aberasi Kromosom pada Tumor Padat Ecie Budiyanti; Des Suryani; Jeanne Adiwinata Pawitan
Majalah Patologi Indonesia Vol 23 No 1 (2014): MPI
Publisher : Perhimpunan Dokter Spesialis Patologi Indonesia (IAPI)

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Abstract

In solid tumor, conventional cytogenetic method usualy fails to detect chromosomal aberation due to failure in obtaining good quality metaphases. Therefore, special technique is required to get the karyotype of solid tumors. Whole chromosome painting (WCP) is a method to paint the chromosomes using Fluorescence In Situ Hybridization (FISH) technique. Application of WCP, both using one and multiple colors (mFISH or SKY) or their modifications can be used to identify chromosomal aberrations, which are valuable in helping to attain a diagnosis, to determine the prognosis and management, and to predict the response to therapy. Key words: chromosomal aberation, FISH, solid tumor, whole chromosome painting.
Sel Punca Kanker (Cancer Stem Cells): Sebuah tinjauan pustaka K Kartini; Jeanne Adiwinata Pawitan
Majalah Patologi Indonesia Vol 27 No 3 (2018): MPI
Publisher : Perhimpunan Dokter Spesialis Patologi Indonesia (IAPI)

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Abstract

Kanker merupakan jaringan yang mengalami penyimpangan dalam pertumbuhannya dan kemungkinan terjadi sebagai respons terhadap kerusakan jaringan. Sel punca yang secara normal diperlukan dalam perbaikan jaringan yang rusak, kini dipertimbangkan memiliki peran dalam pembentukan, pertumbuhan, dan pemeliharaan tumor. Para peneliti dari kelompok tertentu menemukan bahwa suatu jenis sel punca dapat menyebabkan kanker. Pembelahan sel punca secara asimetrik merupakan syarat paling mendasar bagi perkembangan organisme multiselular. Kegagalan dalam pembelahan asimetrik dapat memicu terjadinya tumorigenesis. Proses tumorigenesis membutuhkan kondisi tambahan seperti perluasan faktor lingkungan mikro (stem cell niche) yang merupakan faktor ekstrinsik, atau mutasi sel punca sehingga tidak lagi bergantung pada niche. Sel penginisiasi kanker dikenal dengan sel punca kanker (cancer stem cells). Sel punca kanker (SPK) saat ini dilaporkan banyak ditemukan pada tumor manusia dengan menggunakan metode identifikasi yang umumnya digunakan pada sel punca normal. Penemuan tentang sel punca kanker dan karakteristiknya membawa suatu harapan baru dalam menentukan prognosis kekambuhan dan terapi kanker. Penemuan terapi anti kanker baru yang target spesifiknya adalah sel kanker yang undifferentiated dan dormant dengan dipandu oleh petanda gen SPK dalam proses terapinya, diharapkan meningkatkan angka perbaikan dalam pengobatan pasien kanker.
Peningkatan Kadar Imunosupresan IDO (Indoleamin 2,3-dioksigenase) pada Supernatan Kultur Sel Punca Mesenkim yang Distimulasi dengan Agregat Imunoglobulin G DIAN RATIH LAKSMITAWATI; JEANNE ADIWINATA PAWITAN; MOHAMAD SADIKIN; CAROLINE TAN SARDJONO
JURNAL ILMU KEFARMASIAN INDONESIA Vol 12 No 1 (2014): JIFI
Publisher : Fakultas Farmasi Universitas Pancasila

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Abstract

Mesenchymal stem cells have been known to have the nature of supressing immune response. This cell have been proven to reduce the symptoms of organ transplants rejection. Clinically proofing is also being performed on an autoimmune disease caused by the complex immune such as systemic lupus erythematosus (SLE). This study aimed to know aggregated IgG-stimulating effects of mesenchymal stem cells in relation to IDO secretion, an immunosuppressant agent. Mesenchymal stem cells was isolated from liposuction waste and characterized based on its surface molecule accrording to ISCT criterias. The confirmed cell culture was used to the next culture by the addition of heat aggragated immunoglobulins gamma (HAGG) for six days. IDO secreted on supernatant culture was collected and assayed by elisa method at 450 nm. The results showed that there was elevated levels of IDO (16.73-49.5%) on culture with HAGG stimulation than without stimulation. This study could be a part of the basic
CD34+ UCB stem cells attenuate TGF-β signaling and inhibit liver fibrosis: A new avenue for liver cirrhosis-carcinogenesis prevention Septiana, Wahyunia Likhayati; Antarianto, Radiana Dhewayani; Louisa, Melva; Jusuf, Ahmad Aulia; Barasila, Atikah Chalida; Pawitan, Jeanne Adiwinata; Fasha, Iqbal
Makara Journal of Health Research Vol. 24, No. 2
Publisher : UI Scholars Hub

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Abstract

Background: The liver microenvironment plays a key role in liver fibrosis and carcinogenesis. This study aimed to fill the gap in knowledge on the interaction between hepatic stellate cells and endothelial progenitor cells with biomarkers of liver fibrosis and/or carcinogenesis, including Col1A1, TGF-β, and tenascin-C. Methods: CD34+ stem cells were isolated from umbilical-cord-blood mononuclear cells. 2D and 3D co-culture of CD34+ UCB SCs and LX2 was performed. The cells were incubated in a CO2 incubator for three days. Morphological observation, qRT-PCR of TGF-β1 and COL1A1, and immunocytochemistry of tenascin-C were performed. Results: CD34+ UCB SCs were viable in the 2D and 3D co-culture for 24 h. 3D co-culture of CD34+ UCB SCs and LX2 inhibited in vitro liver fibrosis by lowering Col 1A1 expression as compared to control. We observed lower TGF-β expression in 3D co-culture on days 1 and 2 followed by higher expression of TGF-β on day 3. 2D co-culture of CD34+ UCB SCs and LX2 showed a different level of COL1A1 and TGF- β expression compared with 3D co-culture. Spheroids from 2D co-culture of CD34+ UCB SCs and LX-2 showed immunoreactivity against tenascin-C. Conclusion: Interaction between LX-2 and CD34+ UCB SCs in 3D co-culture inhibits in vitro liver fibrosis. The viability of CD34+ UCB SCs is essential for attenuation of TGF-β signaling in LX-2.
Sel Punca Kanker Kolorektal Noza Hilbertina; Jeanne Adiwinata Pawitan
Cerdika: Jurnal Ilmiah Indonesia Vol. 1 No. 4 (2021): Cerdika : Jurnal ilmiah Indonesia
Publisher : Publikasi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (306.619 KB) | DOI: 10.59141/cerdika.v1i4.70

Abstract

Latar belakang: Kemoterapi dan radioterapi pasien kanker kolorektal sering gagal, atau kambuh kembali, yang disebabkan adanya kelompokan sel dengan sifat sel punca yang disebut sel punca kanker kolorektal. Tujuan penelitian ini adalah mengetahui asal sel punca kolorektal dan sifatnya. Metode penelitian: Untuk menulis studi pustaka ini kami melakukan pencarian di Pubmed/ Medline dan Google Scholar dan memilih artikel yang berkaitan dengan sel punca kanker kolorektal, dalam hal asalnya, cara isolasi dan identifikasinya dan dampak sel punca kanker kolorektal pada prognosis kanker kolorektal. Data dianalisis, dikelompokkan dalam subjudul, dibuat tabel dan ditampilkan secara naratif. Hasil dan pembahasan: Isolasi dan identifikasi sel punca kanker kolorektal dapat dilakukan melalui berbagai sifatnya seperti kemampuan membentuk spheres, dye exclusion dan keberadaan marka permukaan sel. Selain itu, identifikasi dapat dilakukan dengan melihat jalur pensinyalan, aktifitas enzim, marka faktor transkripsi dan uji invivo sebagai baku emas. Kesimpulan: Sel punca kanker kolorektal diduga berasal dari sel punca kolon yang mengalami mutasi dan berhubungan dengan kemampuan invasi, metastasis dan juga survival penderita kanker tersebut.
Prospect of umbilical cord mesenchymal stem cell culture waste in regenerative medicine Jeanne Adiwinata Pawitan; Melisa Leviana; Dewi Sukmawati; Isabella Kurnia Liem; Ria Margiana; Twidy Tarcisia
Journal of Global Pharma Technology Volume 9 Issue 07
Publisher : Journal of Global Pharma Technology

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Abstract

Abstract: Objective:to analyzevarious growth factor (GF) content in umbilical cord derived mesenchymal stem cell (UC-MSC) culture waste at harvest.Methods: We cultured UC- MSCs in complete medium, and collected the medium waste at harvests.  We noted the time of harvests, and measured the concentrations of EGF, VEGF, NGF and Pl GF in the various medium wastes using ELISA method. We grouped GF concentrations according to various culture lengthsat harvest, and calculated themean values and standard deviations of the various GF. We compareddescriptively the GF concentrations at various culture lengths at harvests to conclude which condition yielded the overall highest GF concentration.Results: EGF, VEGF and PlGF concentrations were highest at harvest day-3, whileNGF concentration was the highest at harvest-day-5, respectively. Therefore, overall highest concentration of most GF was at harvest day 3.Conclusion:UC-MSC culture waste at harvest day-3 containedEGF, VEGF, NGF and PlGF, which might be used in regenerative medicine.
PASSAGE INFLUENCE ON THE AGING PROFILE AFTER SUCCESSIVE CRYOPRESERVATION OF BONE MARROW MESENCHYMAL STEM CELLS Ismail Hadisoebroto Dilogo; Starifulkani Arif; Jeanne Adiwinata Pawitan; Ria Anggraeni; Yogi Ismail Gani
Journal of Global Pharma Technology Volume. 9 Issue 6
Publisher : Journal of Global Pharma Technology

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Abstract

Objective: Bone marrow mesenchymal stem cells are promising in cell therapy but the low frequency of this subpopulation necessitates their in vitro expansion and cryopreservation prior to clinical use. The aim of this study was to analyze the passage effect on the aging profile of previously successive cryopreserved bone marrow mesenchymal stem cells.  Methods: The samples of this in-vitro observational analytic study (April - September 2016) were iliac crest bone marrow mesenchymal stem cells that were cryopreserved in passage one and a second group was cryopreserved after being subcultured. It was obtained from UPT TK Sel Punca RSCM-FKUI. Samples were analyzed for each passage in terms of cell size, viability, population doubling time (PDT), and percentage of senescent cells by ANOVA test and independent t-test for the inter-cryopreservation group.Results: The remarkable manifestation of senescence appeared at passage six in both cryopreserved groups, but tend to appear earlier in the twice cryopreservation group. The senescent positive cell size became bigger along with the increasing number of passage. There were significant differences in PDT, 30% confluent cell size, viability, in passage six between the two cryopreservation groups (p<0.001, p<0.001, p=0.022), respectively, but no significant difference in percentage of senescent cells (p= 0.052). The cells became senescent in P6 in both groups (P=0.000).Conclusion: The onset of senescence of bone marrow mesenchymal stem cells was in passage six, while PDT, cell size and viability differ significantly due to successive cryopreservation.
The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast Melinda Remelia; Budiman Bela; Silvia Tri Widyaningtyas; Radiana Dhewayani Antarianto; Nuzli Fahdia Mazfufah; Jeanne Adiwinata Pawitan
Molecular and Cellular Biomedical Sciences Vol 7, No 1 (2023)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v7i1.279

Abstract

Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibroblast transfection to avoid the malignancy issue.Materials and methods: The constructions of CCAT/enhancer binding protein alpha (CEBPA), hepatocyte nuclear factor 4 alpha (HNF4A), nuclear receptor subfamily 1 group I member 2 (NR1I2) were confirmed with DNA digestion and sequencing. Breast reduction (BRED) and palate (PAL) tissue were used as human primary fibroblast sources. The transcription factors were delivered into BRED and PAL with recombination of avian leukosis sarcoma virus (ALSV), human immunodeficiency virus (HIV) matrix, and regulator of expression of virion proteins (Rev) (ALMR), tagged with enhanced green fluorescence protein (eGFP). Post-transfection cells were then cultivated with optimized medium. Gene expression was measured with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).Results: Gene expression levels of CEBPA, HNF4A, NR1I2, glutamate-ammonia ligase (GLUL), albumin (ALB), and cytochrome P450 (CYP) were increased. Transfection with ALMR, which were more efficient in BRED than PAL fibroblasts may have the advantage in autologous cell therapy for elderly patients.Conclusion: Transfection of transcription factors to human primary fibroblast may be performed by using constructions of plasmid as designed in this study.Keywords: recombinant plasmid, hepatocyte-like cells, primary fibroblasts, recombinant peptide, cell reprogramming, autologous cells therapy
Pengaruh Penggantian Medium terhadap Viabilitas Hepatosit Kultur 3D Organoid Hati Sibuea, Christine Verawaty; Pawitan, Jeanne Adiwinata; Antarianto, Radiana
Nommensen Journal of Medicine Vol 7 No 2 (2022): Nommensen Journal of Medicine: Edisi Februari
Publisher : Universitas HKBP Nommensen

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (59.803 KB) | DOI: 10.36655/njm.v7i2.625

Abstract

Background : Liver organoids can be used as materials for Bioartificial Liver, to study the mechanism of liver disease and as drug test toxicity. Reconstruction of liver organoids requires optimal culture methods, culture medium and cellular components to construct liver organoids that resemble liver microstructure in vivo with optimal function. 3D culture method using hepatocytes and stem cells with PRP supplemented William's E can reconstruct liver organoids with liver function. Medium exchange is an usual method to maintain the required nutrients and to eliminate waste products, but it requires a sufficient supply of medium and supplementation. Method and the use of effective and efficient medium with optimal hepatocyte viability are urgently needed in the reconstruction of liver organoids. Objective : This study was aimed to compare the viability of primary hepatocytes in culture medium exchange liver organoids and monoculture and without culture medium exchange. Methods : Primary hepatocytes isolated from Sprague Dawley-Rats mice (250gr, n=3) were co-cultured with umbilical cord mesenchymal stem cells, cord blood CD34+ stem cells and LX2 in PRP-supplemented William's E for 14 days. The culture medium was exchanged at 48 hours, day 7 and day 14 and no culture medium exchanged in the control group. Hepatocyte viability was analyzed using the Trypan Blue Exclusion Test at 48 hours, day 7 and day 14. Results : Hepatocyte viability in culture medium exchange liver organoids was higher than without culture medium exchange, especially in monoculture, but there was no significant difference (p value> 0.05). Conclusion: Hepatocyte viability in culture medium exchange liver organoids was not significantly different from no culture medium exchange liver organoids. Culture medium exchange in monoculture supported hepatocyte viability up to day 14. Keywords: hepatocytes, liver organoids, viability, culture medium ABSTRAK Latar belakang : Organoid hati dapat digunakan sebagai bahan Bioartificial Liver, mempelajari mekanisme penyakit hati dan uji toksisitas obat. Rekonstruksi organoid hati membutuhkan metode kultur, medium kultur dan komponen seluler yang optimal untuk menghasilkan organoid hati yang menyerupai mikrostruktur hati in vivo dengan fungsi yang optimal. Metode kultur 3D menggunakan hepatosit dan sel punca mesenkimal dengan William’s E yang disuplementasi PRP dapat merekonstruksi organoid hati dengan fungsi hati. Pergantian medium merupakan metode yang sering dilakukan untuk mempertahankan nutrisi yang dibutuhkan dan untuk membuang sisa metabolit sel, tetapi membutuhkan persediaan medium dan suplementasi yang cukup banyak. Metode dan penggunaan medium yang efektif dan efisien dengan viabilitas hepatosit yang optimal sangat dibutuhkan dalam rekonstruksi organoid hati. Tujuan : Penelitian ini bertujuan untuk mengetahui perbandingan viabilitas hepatosit primer pada organoid hati dengan pergantian medium kultur dan tanpa pergantian medium kultur. Metode : Hepatosit primer yang diisolasi dari tikus Sprague Dawley-Rats (250gr, n=3) diko-kultur dengan sel punca mesenkimal asal tali pusat, sel punca CD34+ asal darah tali pusat dan LX2 dalam William’s E yang disuplementasi PRP selama 14 hari. Medium kultur diganti pada 48 jam, hari ke-7 dan hari ke-14 dan tidak dilakukan pergantian medium pada kelompok kontrol. Viabilitas hepatosit dianalisa dengan menggunakan Trypan Blue Exclusion Test pada 48 jam, hari ke-7 dan hari ke-14. Hasil : Viabilitas hepatosit pada organoid hati dengan pergantian medium kultur tampak lebih banyak dibandingkan tanpa pergantian medium kultur khususnya pada monokultur, tetapi tidak terdapat perbedaan yang signifikan (nilai p>0,05). Kesimpulan : Viabilitas hepatosit pada organoid hati dengan pergantian medium kultur tidak berbeda secara signifikan dengan organoid hati tanpa pergantian medium kultur. Pergantian medium kultur pada monokultur mendukung viabilitas hepatosit hingga hari ke-14. Kata Kunci : Hepatosit, organoid hati, viabilitas, medium kultur
ADIPOSE-DERIVED STEM CELL THERAPY ON NON-COMMUNICABLE DISEASE: A SYSTEMATIC REVIEW Tandarto, Kevin; Purwoko, Reza Yuridian; Oktarina, Caroline; Jonlean, Reganedgary; Irawan, Cosphiadi; Abdullah, Murdani; Pawitan , Jeanne Adiwinata
Journal of Stem Cell Research and Tissue Engineering Vol. 7 No. 1 (2023): JOURNAL OF STEM CELL RESEARCH AND TISSUE ENGINEERING
Publisher : Stem Cell Research and Development Center, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jscrte.v7i1.40636

Abstract

The increasing number of non-communicable diseases demands practical therapy innovations, including adipose-derived stem cell application. This study aimed to analyze the effectiveness of adipose stem cell therapy on non-communicable disease patients. The method used in this study was a systematic review according to PRISMA 2020 guidelines. The database search was done on PubMed, Google Scholar, Proquest, and the EBSCO host database between 2016 and 2021. ROBINS-I tool and RoB-2 were used to assess the risk of bias in the clinical trial study. The first literature search identified a total of 2615 articles. After exclusion for some reason, 6 articles were included in this systematic review study. A total of five studies were included in this study. Based on the risk of bias assessment of the included studies, it was found that all studies had a low risk of bias in all domains. This study showed that the efficacy of adipose-derived stem cell therapy was inconsistent; however, the results were promising. In addition, the results showed that adipose-derived stem cell therapy was safe without significant side effects. Further study was needed to identify therapeutic strategies based on Evidence-based Medicine (EBM).