scholarly journals KONDROGENESIS ADIPOSE-DERIVED STEM CELLS MENGGUNAKAN PLATELET-RICH PLASMA PADA SCAFFOLD SUTRA

2020 ◽  
Vol 13 (1) ◽  
pp. 31-38
Author(s):  
Imam Rosadi ◽  
Karina Karina ◽  
Komang A. Wahyuningsih ◽  
Iis Rosliana ◽  
Tias Widyastuti ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bombyx Mori ◽  
Silk Fibroin ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Silk Fibroin Scaffold ◽  
Salt Leaching ◽  
Fetal Bovine

AbstrakStudi tentang kemampuan adipose-derived stem cells (ADSCs) sebagai sel punca yang dapat berdiferensiasi menjadi kondrosit menggunakan platelet-rich plasma (PRP) sebagai subtitusi fetal bovine serum (FBS) telah banyak dilaporkan. Penggunaan medium pertumbuhan dengan kombinasi ADSCs, PRP dan scaffold sutra masih belum banyak dipelajari dalam rekayasa jaringan kartilago. Studi ini bertujuan untuk mempelajari pengaruh medium yang mengandung 5%, 10% dan 20% PRP terhadap proses kondrogenesis ADSCs manusia yang dikultur pada scaffold sutra Bombyx mori Indonesia. Metode penelitian diawali dengan pembuatan scaffold sutra menggunakan metode salt-leaching, isolasi dan kultur ADSCs manusia dari jaringan lemak, uji pertumbuhan ADSCs pada scaffold sutra dengan variasi konsentrasi PRP pada medium serta analisis kadar glikosaminoglikan (GAG). Hasil penelitian menunjukkan bahwa ADSCs yang dikultur menggunakan PRP lebih tinggi laju pertumbuhannya dibandingkan dikultur menggunakan FBS selama 7 hari pengamatan. Kadar GAG yang disekresikan ADSCs kelompok PRP juga lebih tinggi dibandingkan kelompok FBS. Kadar GAG tertinggi pada hari ke-21 pengamatan adalah medium yang mengandung 20% PRP kemudian 10% dan 5%, sedangkan kadar GAG kelompok kontrol cenderung stabil pada kadar yang rendah. Berdasarkan hasil tersebut, medium yang mengandung PRP memiliki potensi dalam menginduksi kondrogenesis ADSCs yang dikultur pada scaffold sutra.Abstract The studies on adipose-derived stem cells (ADSCs) differentiation into chondrocytes using platelet-rich plasma (PRP) as a substitute for fetal bovine serum (FBS) have been reported. However, the combination of ADSCs, PRP and silk fibroin scaffold has not been widely studied for developing cartilage engineering. Therefore, this research aims to study the effect of medium containing 5%, 10% and 20% PRP towards chondrogenesis of human ADSCs cultured on silk fibroin scaffold from Indonesia Bombyx mori. At first, the silk fibroin scaffold was fabricated using a salt-leaching method, then ADSCs were isolated and cultured from adipose tissues. The assays of growth curve and biocompatibility of silk fibroin scaffold toward ADSCs supplemented by PRP as well as glycosaminoglycans (GAG) concentration were conducted later. The results showed that higher absorbance of proliferation rate was on ADSCs supplemented by various PRP concentrations compare to FBS control group for seven days of observation. Level of GAG, which secreted by ADSCs supplemented by a various concentration of PRP, was also higher than the FBS group. The highest level of GAG on day 21 was observed in 20% PRP group then 10% and 5% PRP, while a group of GAG level is stable at low levels. This study concludes that PRP has the potential to induce chondrogenesis ADSCs which cultured on silk fibroin scaffold.

scholarly journals Biokompatibilitas scaffold sutera asal Bombyx mori ukuran pori 100µm terhadap adipose-derived stem cells (ADSCs) yang dikultur pada berbagai medium pertumbuhan

2020 ◽  
Vol 24 (1) ◽  
pp. 7
Author(s):  
Imam Rosadi ◽  
Karina Karina ◽  
Komang A. Wahyuningsih ◽  
Anggraini Barlian ◽  
Iis Rosliana ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ascorbic Acid ◽  
Bombyx Mori ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Fetal Bovine

Rekayasa jaringan terdiri dari 3 komponen utama yaitu sel, nutrisi, dan scaffold. Penggunaan sel punca asal jaringan lemak (adipose-derived stem cells/ ADSCs) telah banyak dikembangkan sebagai sumber sel dalam teknologi rekayasa jaringan. Medium yang digunakan dalam mendukung pertumbuhan sel diantaranya medium yang mengandung serum seperti fetal bovine serum (FBS), kombinasi FBS dan L-ascorbic acid 2-phosphate (LAA) atau platelet-rich plasma (PRP). Pada penelitian ini, sutera asal Bombyx mori diproduksi menjadi scaffold sutera ukuran pori 100µm kemudian ADSCs dikultur diatas scaffold dalam medium mengandung 10% FBS, 10% FBS-LAA atau 10% PRP. Uji yang dilakukan adalah uji pertumbuhan ADSCs yang dikultur pada polystyrene kemudian uji biokompabilitas scaffold sutera pada ADSCs dalam medium mengandung 10% FBS, 10% FBS-LAA dan 10% PRP. Hasilnya menunjukkan bahwa ketiga kelompok ADSCs dalam variasi medium yang mengandung FBS, FBS-LAA atau PRP dapat mendukung pertumbuhan sel. Ketiga medium tersebut juga tidak berbeda bermakna antar kelompok pada uji biokompabilitas ADSCs yang dikultur pada scaffold sutera. Berdasarkan hasil tersebut, scaffold sutera berpotensi sebagai substrat ADSCs yang dapat dikembangkan sebagai teknologi rekayasa jaringan.

scholarly journals Perbandingan Morfologi Adipose-Derived Stem Cells asal Donor Diabetes Melitus Tipe 2 dalam Medium Mengandung Platelet-Rich Plasma dan Fetal Bovine Serum

2019 ◽  
Vol 1 (2) ◽  
pp. 64
Author(s):  
Karina Karina ◽  
Imam Rosadi ◽  
Siti Sobariah ◽  
Iis Rosliana ◽  
Komang A Wahyuningsih ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Fetal Bovine ◽  
Diabetes Melitus

Salah satu terapi luka pada diabetes melitus tipe 2 adalah terapi sel punca. Lingkungan mikro bagi sel termasuk sel punca, dapat rusak akibat komplikasi dari diabetes. Lingkungan mikro yang rusak tersebut dapat menyebabkan penuaan (senescent) dini pada sel punca. Studi ini bertujuan untuk menganalisis pengaruh morfologi adipose-derived stem cells (ADSCs) dari donor diabetes mellitus tipe 2 dalam medium yang mengandung platelet-rich plasma (PRP). Tahapan studi yang dilakukan yaitu menghitung variasi densitas awal ADSCs. Densitas awal untuk kultur ADSCs adalah 5,000; 10,000; dan 20,000 sel pada setiap kelompok. Hasilnya menunjukkan bahwa morfologi ADSCs dalam medium PRP umumnya lebih kecil dibandingkan morfologi ADSCs dalam medium FBS pada berbagai jumlah densitas awal ADSCs yang dikultur. Morfologi ADSCs kelompok PRP didapatkan semakin kecil luas morfologinya pada densitas kultur awal sel yang besar (20,000 = 0,014 mm2; 10,000 = 0,016 mm2; 5,000 = 0,018 mm2) begitu juga kelompok FBS (20,000 = 0,032 mm2; 10,000 = 0,032 mm2; 5,000 = 0,036 mm2). Luas ukuran ADSCs yang dikultur menggunakan FBS jauh lebih besar dibandingkan PRP dan berbeda bermakna pada densitas jumlah sel awal yang dikultur sebanyak 20,000 sel (p <0,05). Berdasarkan hasil tersebut menunjukkan bahwa morfologi ADSCs yang dikultur dengan kerapatan densitas 20,000 sel dalam medium PRP memiliki ukuran sel yang lebih kecil secara signifikan dibandingkan medium FBS.

scholarly journals Differentiation of adipose-derived stem cells into Schwann-like cells: fetal bovine serum or human serum?

2015 ◽  
Vol 48 (3) ◽  
pp. 170 ◽  
Author(s):  
Elham Younesi ◽  
Vahid Bayati ◽  
Mahmoud Hashemitabar ◽  
Seyyed Saeed Azandeh ◽  
Dariush Bijannejad ◽  
...  
Keyword(s):  
Stem Cells ◽  
Human Serum ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Adipose Derived Stem Cells ◽  
Fetal Bovine

scholarly journals Role of Hypoxia on Growth and Differentiation of Human Adipose Derived Stem Cells Grown on Silk Fibroin Scaffold Induced by Platelet Rich Plasma

2021 ◽  
Vol 53 (3) ◽  
pp. 415-427
Author(s):  
Anggraini Barlian ◽  
Marselina Irasonia Tan ◽  
Ergha Widya Sarjana ◽  
Noviana Vanawati
Keyword(s):  
Stem Cells ◽  
Silk Fibroin ◽  
Platelet Rich Plasma ◽  
Adipose Derived Stem Cells ◽  
Low Oxygen ◽  
Silk Fibroin Scaffold ◽  
Hypoxic Conditions ◽  
Proliferation And Differentiation ◽  
The Difference ◽  
Low Oxygen Concentration

Previous research has proven that 10% platelet-rich plasma (PRP) can enhance growth and differentiation of human adipose derived stem cells (hADSC) grown on silk fibroin scaffold into chondrocytes. A low oxygen concentration (hypoxia) condition is an important factor that potentially affects the ability of hADSC to grow and differentiate. The objective of this research was to analyze the difference in growth and differentiation capacity of hADSC grown on salt leached silk fibroin scaffold supplemented by 10% PRP under normoxic and hypoxic conditions. The growth capacity of the hADSC was determined by MTT assay and differentiation was tested using glycosaminoglycan (GAG) content analysis, while chondrocyte markers were visualized with the immunocytochemistry (ICC) method. This research observed hADSC proliferation under normoxic and hypoxic conditions for 21 days. Visualization of type 2 collagen showed that it was more abundant under hypoxia compared to normoxia.  HIF-1α was only detected in the hADSC cultured in hypoxic conditions. In conclusion, culture under hypoxic conditions increases the capacity of hADSC to grow and differentiate into chondrocytes. This is the first study that has shown that hypoxia is able to enhance the proliferation and differentiation of hADSC grown on 3D salt leached silk fibroin scaffold supplemented by 10% PRP.

Fetal bovine serum-free cryopreservation methods for clinical banking of human adipose-derived stem cells

Cryobiology ◽  
2018 ◽  
Vol 81 ◽  
pp. 65-73 ◽  
Author(s):  
Seah Park ◽  
Dong Ryul Lee ◽  
Ji Sun Nam ◽  
Chul Woo Ahn ◽  
Haekwon Kim
Keyword(s):  
Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Adipose Derived Stem Cells ◽  
Serum Free ◽  
Fetal Bovine

The effect of activated pure platelete rich plasma (P-PRP) on the proliferation of adipose -derived mesenchymal stem cells (AD-MSCs) as a substitute for fetal bovine serum (FBS)

2019 ◽  
Vol 20 (1) ◽  
pp. 35-40
Author(s):  
Aya El-Gamal
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Risk Of Infection ◽  
Self Renewal ◽  
Infection Transmission ◽  
Fetal Bovine ◽  
Subsequent Risk

The field of stem cells has attracted the attention of many researchers as a hope to treat many incurable diseases because of their ability of self-renewal and differentiation into a specific type of cells that compensates damaged cells. Many studies have been published to confirm their effectiveness but there are some obstacles that limit their clinical applications. One of these obstacles is xenogenic fetal bovine serum (FBS); the main proliferative source for stem cell culture with subsequent risk of infection transmission or immunogenic problems. So, in our study, we aimed to replace the xenogenic FBS with xeno-free blood derivative. We tested the effect of different concentrations of activated pure platelet rich plasma (P-PRP); one of the blood derivatives on proliferation of adipose derived mesenchymal stem cells (AD-MSCs) in comparison to FBS and found that 20% activated P-PRP followed by 10% activated P-PRP increased the proliferation rate of AD-MSCs more than 10% FBS.

scholarly journals Stimulatory Role of Glycated Fetal Bovine Serum Along with Iron on In Vitro Production of Insulin by Differentiated Mouse Embryonic Stem Cells

2011 ◽  
Vol 57 (4) ◽  
pp. 356-361
Author(s):  
Ikuo Nishigaki ◽  
Gowri Rangasamy Gunassekaran ◽  
Panjan Nagappan Venkatesan ◽  
Mandupal Chaco Sabu ◽  
Sabu Priya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Embryonic Stem ◽  
In Vitro Production ◽  
Fetal Bovine ◽  
Vitro Production

Pooled thrombin-activated platelet-rich plasma: a substitute for fetal bovine serum in the engineering of osteogenic/vasculogenic grafts

2015 ◽  
Vol 11 (5) ◽  
pp. 1542-1552 ◽  
Author(s):  
Laurent A. Tchang ◽  
Benjamin E. Pippenger ◽  
Atanas Todorov ◽  
Francine Wolf ◽  
Maximilian G. Burger ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Platelet Rich Plasma ◽  
Fetal Bovine

302 REPLACEMENT OF FETAL BOVINE SERUM WITH KNOCKOUT SERUM REPLACER AND STEM CELLS CONDITIONED MEDIUM DURING IN VITRO CULTURE OF BOVINE EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 307
Author(s):  
M. M. Souza ◽  
N. Z. Saraiva ◽  
C. S. Oliveira ◽  
T. A. D. Tetzner-Nanzeri ◽  
R. Vantini ◽  
...  
Keyword(s):  
Stem Cells ◽  
In Vitro Culture ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Protein Supplementation ◽  
Control Group ◽  
Bovine Embryos ◽  
Fetal Bovine

The use of fetal bovine serum (FBS) as protein supplementation in IVP of bovine embryos has presented difficulties because it can introduce a number of pathogenic components in culture systems, can be related to the birth of calf with abnormal growth and development, and precludes the establishment of the actual nutritional needs of the embryo, because it contains an unlimited variety of substances. This study evaluated the replacement of the FBS in the medium of in vitro culture (IVC) of bovine embryos, using the knockout serum replacer (KSR) as protein supplementation and culture medium conditioned with stem cells. Therefore, bovine oocytes from ovaries of slaughterhouse were selected and matured in vitro in TCM-199 medium supplemented with 10% FBS (Crypion), 1.0 μg mL-1 FSH (Pluset®, Calier, Barcelona, Spain), 50 μg mL-1 hCG (Profasi®, Serono, Geneva, Switzerland), 1.0 μg mL-1 estradiol (Sigma E-2758, Sigma Chemical, St. Louis, MO, USA), 0.2 mM sodium pyruvate, and 83.4 μg mL-1 amikacin for 24 h. After that, 1144 oocytes were fertilized in IVF-TALP medium containing 6 mg mL-1 of BSA. After 18 to 22 h, the zygotes were cultured in SOF + 5% FBS (group 2); SOF + 5% KSR (group 3); SOF (5% FBS) + 10% SOF (5% FBS) conditioned by stem cells (group 4); or SOF (5% KSR) + 10% SOF (5% KSR) conditioned by stem cells (group 5), in an atmosphere of 5% O2 at 38.5°C for 8 days. A control group outside the controlled atmosphere was added, supplemented with 5% FBS (group 1). The SOF medium supplemented with 5% FBS or KSR was conditioned by stem cells and added to SOF medium for the culture of embryo at a concentration of 10%. The rates of cleavage and production of blastocysts were assessed 48 hours and 7 days after IVF, respectively, and analyzed by chi-square test, with a significance level of 5% in the statistical program Minitab® (release 14.1, Minitab, State College, PA, USA). On the eighth day, the TUNEL test for determination of the percentage of apoptosis and the differential staining technique for determination of inner cell mass (ICM) and trophoblast (TF) were performed. The results were submitted to ANOVA, followed by comparing the means by Tukey’s test using the program GraphPad Prism (GraphPad, San Diego, CA, USA). The treatments did not differ in the production of embryos, being similar to the control group: G1 = 31.75% (74/233), G2 = 35.26% (79/224), G3 = 32.70% (74/226), G4 = 28.76% (63/219), and G5 = 26.85% (65/242). With regard to the assessment of embryonic quality, the treatments showed similar results to the control groups. No differences were observed among groups both in color and ICM/TF ratio (G1 = 0.60, G2 = 0.62, G3 =0.65, G4 = 0.60, and G5 = 0.60). Furthermore, the TUNEL showed no significant difference in the percentage of apoptosis among groups (G1 = 7.10%, G2 = 3.76%, G3 = 5.58%, G4 = 4.50%, and G5 = 4.11%). The data obtained so far indicate that it is possible to produce embryos in vitro by replacing the FBS in the culture, achieving results similar to those obtained with serum. Financial support: FAPESP 2007/58506-6.

scholarly journals Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells

Cells ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 724 ◽  
Author(s):  
Mathew Cowper ◽  
Trivia Frazier ◽  
Xiying Wu ◽  
J. Curley ◽  
Michelle Ma ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Human Platelets ◽  
Dependent Manner ◽  
Wide Range ◽  
Human Ascs ◽  
Fetal Bovine ◽  
Stromal Stem Cells

Introduction: Adipose derived stromal/stem cells (ASCs) hold potential as cell therapeutics for a wide range of disease states; however, many expansion protocols rely on the use of fetal bovine serum (FBS) as a cell culture nutrient supplement. The current study explores the substitution of lysates from expired human platelets (HPLs) as an FBS substitute. Methods: Expired human platelets from an authorized blood center were lysed by freeze/thawing and used to examine human ASCs with respect to proliferation using hematocytometer cell counts, colony forming unit-fibroblast (CFU-F) frequency, surface immunophenotype by flow cytometry, and tri-lineage (adipocyte, chondrocyte, osteoblast) differentiation potential by histochemical staining. Results: The proliferation assays demonstrated that HPLs supported ASC proliferation in a concentration dependent manner, reaching levels that exceeded that observed in the presence of 10% FBS. The concentration of 0.75% HPLs was equivalent to 10% FBS when utilized in cell culture media with respect to proliferation, immunophenotype, and CFU-F frequency. When added to osteogenic, adipogenic, and chondrogenic differentiation media, both supplements showed appropriate differentiation by staining. Conclusion: HPLs is an effective substitute for FBS in the culture, expansion and differentiation of human ASCs suitable for pre-clinical studies; however, additional assays and analyses will be necessary to validate HPLs for clinical applications and regulatory approval.

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