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Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum Tjahjono, Yudy; Dewi, Sianty; Novita, Bernadette Dian; Wijaya, Hendy; Putra, Brilliant Dwi; Kuncorojakti, Suryo; Hendriati, Lucia; Jong, FX Himawan Haryanto; Widodo, Teguh; Malonda, Franklin Vincentius
Universa Medicina Vol. 44 No. 1 (2025)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2025.v44.65-72

Abstract

Background Mesenchymal stem cells (MSCs) are valued in regenerative medicine for their multipotency, proliferative capacity, and immunomodulatory properties. Wharton’s jelly-derived MSCs (WJ-MSCs) from the umbilical cord offer a non-invasive, promising source for clinical applications, because easy isolation, lack of ethical concerns, and the presence of both embryonic and adult stem cells have made them a valuable source for use in therapeutic applications and regenerative medicine. This study aimed to optimize WJ-MSC isolation and characterization methods. Methods Human umbilical cords from three healthy donors were collected post-cesarean under strict inclusion criteria. WJ-MSCs were isolated using the explant culture method, with cells adhering to T75 flasks pre-coated with 2% gelatin. Cultures were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% freshly prepared Human AB serum and monitored for 21 days. Flow cytometry (BD FACSAria) was performed at passages 1 and 5 to assess MSC markers CD105, CD73, CD90, and CD44, alongside the exclusion marker CD45. Results WJ-MSCs exhibited fibroblast-like morphology by passage 1 and showed robust proliferation. Flow cytometry revealed high CD44 expression (~60%) at passage 1, while CD105, CD73, and CD90 became prominent by passage 5. CD45 remained low, suggesting minimal hematopoietic contamination. Conclusion This study confirms the feasibility of isolating and expanding WJ-MSCs using DMEM with 10% human AB serum. While consistent cell growth was achieved, the 21-day culture period may require optimization for scalability, including serum concentration, substrate coatings, and oxygen levels. CPJ-MSCs may be preferable for applications demanding rapid expansion and early marker expression.
Uji Aktivitas Analgesik Tablet Asam 4-(Klorometil)Salisilat pada Mencit Putih dengan Metode Writhing Test dan Hotplate Tiffany, Angela; Ribka Abiati, Meidelin; Foe, Kuncoro; Tjahjono, Yudy; Caroline; Esar, Senny Yesery; Hadinugroho, Wuryanto
JFIOnline | Print ISSN 1412-1107 | e-ISSN 2355-696X Vol. 14 No. 1 (2022): Jurnal Farmasi Indonesia
Publisher : Pengurus Pusat Ikatan Apoteker Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (507.738 KB) | DOI: 10.35617/jfionline.v14i1.78

Abstract

Acetylsalicylic acid is an analgesic drug that still popularly used. However, acetylsalicylic acid may result undesirable side effect of gastrointestinal as it can cause gastric ulcer to bleeding. Previous research has succeeded synthesizing new compound derived from salicylic acid, namely 4-(chloromethyl)salicylic acid which has the potential to reduce its side effects with a better pharmacological effects than acetylsalicylic acid. In its development, the formulation of the compound in tablet dosage form has been carried out. However, until now there has not been a pharmacological test to determine whether it still provides a greater effect than acetylsalicylic acid after being formulated. This study is to test the analgesic activity of 4-(chloromethyl)salicylic acid tablets in albino mice using writhing test and hotplate methods. Analgesic activity test using the hotplate method was carried out on albino mice which given the suspension of the tablet sample and the observations were made in a periodic of time. Analgesic activity test using writhing test method was carried out in 0.6% acetic acid-induced albino mice, where the suspension of the tablet was administered 30 minutes before administration of acetic acid. Analgesic activity was assessed by counting the number of writhings, and the results were statistically compared with one-way ANOVA. Tests were also performed on acetylsalicylic acid as a positive control and suspension as a negative control group.  There were significant differences (P<0.05) of the analgesic activity observed among groups. Both methods show that 4-(chloromethyl)salicylic acid tablets provide stronger analgesic activity than acetylsalicylic acid.
Optimized RT-qPCR Detection of Hepatic Lipopolysaccharide-Binding Protein in Diet-Induced Obese Mice Tjahjono, Yudy; Wedharga, I Gede Putu Adhi; Novita, Bernadette Dian; Tahalele, Paul; Wijaya, Hendy; Soediono, Endang Isbandiati; Hendrata, Adi Pramono; Dewi, Sianty; Wijaya, Sumi; Ervina, Martha; Kuncorojakti, Suryo
Jurnal Medik Veteriner Vol. 8 No. 2 (2025): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jmv.vol8.iss2.2025.308-317

Abstract

High-throughput RT-qPCR results on hepatic lipopolysaccharide-binding protein (LBP) expression in obese subjects are essential, as they reveal the endotoxin’s role in the development of obesity and non-communicable disease (NCD). This study aimed to optimize RT-qPCR detection of LBP in diet-induced obese mice. This study primarily focused on addressing high variability through reference gene normalization. A total of six male C57BL/6 mice aged 6 weeks were randomly allocated into two dietary treatments (n = 3), consisting of mice fed with the standard chow diet (SCD group) and mice fed with the high-fat and high sucrose diet (HFHS group) ad libitum for 8 weeks. Relative quantification strategies involving the standard 2-ΔΔCt method (calibrator as mean) and the modified 2-ΔΔCt method (calibrator as individual sample-matched biological replicates) were compared in terms of their variability. Obesity was successfully induced in the HFHS treatment group, as indicated by significantly higher body weight, calorie intake, and LBP relative expressions compared to the SCD group. In addition, a sample-specific calibrator approach using the modified 2-ΔΔCt method resulted in lower variability in relative gene expression levels. A modified 2-ΔΔCt method, which utilizes a sample-specific calibrator to counteract sample-specific variability, was successfully employed to address high variability in RT-qPCR results.
Deteksi populasi Cd3+Cd4+Cd25+Foxp3+ T-regulator pada Limpa Mencit Galur Balb/c dan Swiss-webster yang Mudah dan Cepat dengan Metode Flow Cytometry Tjahjono, Yudy; Wijaya, Hendy; Esar, Senny Yesery; Caroline, Caroline; Jafet, Nico; Kusuma, I Made Andika Bara; Chrisantia, Oryza; Bhendy, Maria Theresia Primadewy; Palpialy, Sindi Siska; Soehadi, Shellin; Wilianto, Yufita Ratnasari
Jurnal Farmasi Sains dan Terapan (Journal of Pharmacy Science and Practice) Vol. 9 No. 2 (2022): October
Publisher : Faculty of Pharmacy, Widya Mandala Surabaya Catholic University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33508/jfst.v9i2.4079

Abstract

Sel T-regulator (Treg) memainkan peran kunci dalam mengendalikan autoimunitas, respon alergi, peradangan, dan respons terhadap infeksi. Sel T Cd4+ yang secara konstitutif mengekspresikan Cd25 dan protein forkhead 3 (Foxp3; yaitu, Cd3+Cd4+Cd25+Foxp3+) atau disebut juga sebagai Treg fungsional, telah terbukti memainkan peran utama dalam pemeliharaan toleransi dan homeostasis imun, sehingga deteksi Treg fungsional sangat penting dilakukan sebagai salah satu parameter imunomodulasi. Penelitian ini bertujuan untuk mendeteksi populasi sel Treg (Cd3+Cd4+Cd25+Foxp3+) pada organ limpa mencit galur Balb/c dan Swiss-webster dengan menggunakan metode yang mudah, cepat, serta terjangkau. Metode deteksi sel Treg dilakukan dengan cara isolasi organ limpa mencit Balb/c dan Swiss-webster kemudian dilakukan preparasi dan deteksi sel menggunakan instrumen flow cytometry. Hasil populasi sel Treg yang didapatkan berturut-turut pada mencit Balb/c dan Swiss-webster adalah 2,02 ± 0,36% dan 3,14 ± 1,64% dari total populasi yang dideteksi dengan hanya membutuhkan waktu 145 menit sejak pembedahan. Berdasarkan hasil tersebut dapat disimpulkan bahwa pengamatan populasi sel Treg pada splenosit mencit galur Balb/c dan Swiss-webster dengan flow cytometry dapat dilakukan dengan mudah dan praktis dengan pewarnaan kombinasi antibodi yang tertera pada manuskrip ini. Selain itu, metode pewarnaan yang tertera dapat digunakan sebagai salah satu acuan untuk mendeteksi imunomodulator pada berbagai model hewan coba yang menggunakan mencit galur Balb/c dan Swiss-webster, khususnya yang berhubungan dengan populasi sel Treg.