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Latar Belakang: Diabetes melitus (DM) tipe 2 adalah suatu penyakit metabolik yang kompleks dan kronis yang ditandai dengan gangguan angiogenesis. Inflamasi kronis derajat ringan dan stres oksidatif yang meningkat pada DM tipe 2 diketahui dapat menyebabkan gangguan fungsi biologis pada sel progenitor/sel punca vaskular, salah satunya adalah adipose-derived mesenchymal stem cell (ADSC). Sejumlah penelitian menunjukkan potensi vaskulogenik ADSC dan perannya pada regenerasi jaringan. Hingga saat ini aplikasi sel punca autologus pada penderita DM untuk menginisiasi vaskularisasi masih mengalami kendala. Platelet-rich plasma (PRP) diketahui kaya akan berbagai faktor pertumbuhan, termasuk VEGF, yang penting untuk proses angiogenesis.

Tujuan: Penelitian ini bertujuan untuk menguji dan menganalisis efek pemberian PRP PMI terhadap proliferasi (jumlah sel stromal, nilai population doubling time (PDT), dan persentase sel hidup), diferensiasi (pembentukan koloni, ekspresi CD73, CD90, CD105, dan tiga lini diferensiasi), ekspresi mRNA VEGF dan VEGFR2, dan potensi angiogenik ADSC DM in vitro (sekresi VEGF dan pembentukan tubular kapiler).

Metode: Terlebih dahulu, konsentrasi trombosit per µL dan kadar VEGF per 1x10³ trombosit pecah yang terkandung dalam PRP Palang Merah Indonesia (PMI) dibandingkan dengan PRP DM dan non-DM. Lalu, stromal vascular fraction (SVF) diisolasi dari jaringan lemak menggunakan metode enzimatik, dan SVF penderita DM tipe 2 (n= 15) dan non-DM (n= 10)  dikultur dalam medium kontrol hingga didapat ADSC pasase 1−3 (P1−P3). Proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2, serta potensi angiogenik ADSC DM dan non-DM diukur dan dibandingkan. ADSC DM P3 kemudian dikultur dalam medium PRP PMI 5%, 10%, 15%, dan 20%, lalu proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2 diukur dan dibandingkan dengan kontrol (FBS) untuk mendapatkan konsentrasi PRP optimum. ADSC DM P3 yang diprekondisikan dengan PRP optimum, dengan atau tanpa anti-VEGF (bevacizumab) 100 ng/mL, dan ADSC DM P3 kontrol dikultur, lalu sekresi VEGF dan pembentukan tubular kapiler pada Matrigel^® diukur.

Hasil: Pada penelitian ini tidak ditemukan perbedaan bermakna antara konsentrasi trombosit per µL PRP DM, non-DM, dan PMI (p= 0,22). Namun, PRP non-DM memiliki kadar VEGF per 1000 trombosit pecah lebih rendah bermakna (0,20 (0,04−0,35) fg) dibandingkan PRP DM (0,69 (0,21−1,17) fg), p= 0,03) dan PMI (1,84 (1,38−2,10) p= 0,01), dan tidak ada perbedaan bermakna antara PRP DM dan PRP PMI (p= 0,06). Jumlah sel stromal per gram lemak dan jumlah koloni sel stromal DM lebih rendah dari non-DM (86,35 (52,48−106,76) x 10⁶ vs 158,93 (101,59−185,94) x 10⁶, p= 0,01, dan 94 ± 14 koloni vs 31 ± 32 koloni, p= 0,004). Tidak terdapat perbedaan bermakna antara DM dan non-DM pada persentase sel stromal hidup (p= 0,24), ekspresi CD73 (p= 0,21), CD90 (p= 0,90), adipogenesis, kondrogenesis, osteogenesis, PDT P2 (p= 0,27), PDT P3 (p= 0,21), dan persentase sel hidup ADSC P2 (p= 0,07), sedangkan ekspresi CD105 (64,41 (51,20−73,38)% vs 91,40 (82,62−95,47)%, p< 0,001) ADSC DM P1 dan persentase sel hidup (82,70 ± 8,07% vs 91,15 ± 3,77%, p= 0,04) ADSC DM P3 lebih rendah bermakna dibandingkan ADSC non-DM. Tidak ada penurunan yang bermakna pada ekspresi relatif mRNA VEGF (0,64 (0,30−1,08), p= 0,86) dan VEGFR2 DM (0,64 ± 0,56, p= 0,49) jika dibandingkan dengan ADSC non-DM. Rerata kadar VEGF dalam conditioned medium (CM) yang disekresikan oleh 1x10³ ADSC DM dan non-DM secara berturut-turut sebesar 0,74 pg/mL dan 0,62 pg/mL. ADSC DM yang diberi PRP optimum, yaitu 15% memiliki nilai PDT yang lebih rendah (2,33 ± 0,56 hari vs 5,04 ± 1,26 hari, p= 0,01) dan persentase sel hidup (95,53 ± 1,60% vs 78,95 ± 10,13%, p=0,01) yang lebih tinggi bermakna dibandingkan dengan kontrol. Terjadi peningkatan ekspresi CD105, mRNA VEGF, dan VEGFR2 ADSC DM yang diberi PRP 15% (secara berturut-turut 1,81 ± 0,73, p= 0,01; 5,27 ± 5,69, p= 0,23; dan 9,01 ± 11,59, p= 0,06) relatif terhadap kontrol. ADSC DM yang diberi PRP 15% dan ADSC DM kontrol secara berturut-turut mensekresikan VEGF rerata sebanyak 0,57 pg/mL dan 1,67 pg/mL per 1x10³ sel hidup. Jumlah tubular kapiler in vitro ADSC DM yang diberi PRP meningkat pada jam ke-24 jika dibandingkan dengan kontrol dan tidak berbeda bermakna dengan ADSC non-DM, namun membutuhkan waktu lebih panjang, serta tidak berbeda bermakna dengan ADSC DM yang diberi PRP dan anti-VEGF (p=0,78).

Kesimpulan: ADSC DM terbukti mengalami kerusakan selular yang dicirikan dengan penurunan proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2, serta potensi angiogeniknya. Pemberian PRP 15% (VEGF 98,00 pg/mL) dapat memperbaiki kerusakan tersebut melalui efek sinergis yang dihasilkan oleh VEGF dan faktor pertumbuhan lainnya yang terdapat dalam PRP.

 

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Background: Type II diabetes mellitus (DM type 2) is a chronic and complex metabolic disease identified by impaired angiogenesis. Low grade chronic inflammation and increasing oxidative stress in DM type 2 decrease the biological functions of progenitor/stem cells, including adipose-derived stem cells (ADSC).  ADSC plays significant roles in angiogenesis and tissue regeneration. Some studies have shown the vasculogenic potency of ADSC and its role in tissue regeneration. To date, autologous cell application in DM patients to initiate vascularization is hindered. Platelet-rich plasma (PRP) is widely known to contain generous amount of growth factors including VEGF with significant role in angiogenesis.

Objective: This study aimed to investigate and analyze the effect of PRP preconditioning to the proliferation (stromal cell number, population doubling time (PDT) and percentage of viable cells), differentiation (colony formation, CD73, CD90, CD105 expressions, three lineage of differentiation), expression of mRNA VEGF and VEGFR2, as well as in vitro angiogenic potency of ADSC DM (VEGF secretion and capillary tube formation).

Methods: Initially, platelet concentration per µL and VEGF per 1x10³ lysed platelet contained in Palang Merah Indonesia (PMI) PRP was compared to DM and non-DM PRP. Subsequently, stromal vascular fraction (SVF) from 15 DM and 10 non-DM donors was enzymatically isolated from adipose tissue, and cultured in control media to generate passage 1−3 (P1−P3) ADSC. Proliferation, differentiation, mRNA VEGF and VEGFR2 expression, as well as angiogenic potency of DM ADSC in vitro were measured and compared to non-DM control. P3 DM ADSC was then cultured in media contained 5%, 10%, 15%, and 20% PMI PRP, and proliferation, differentiation, mRNA VEGF and VEGFR2 expression were measured and compared to FBS control to determine optimum PMI PRP concentration. P3 DM ADSC preconditioned with optimum PMI PRP, with or without anti-VEGF (bevacizumab) 100 ng/mL, and control DM ADSC were cultured, and VEGF secretion was measured, as well as capillary tube formation on Matrigel^®.

Results: In this study no significant differences were observed between platelet concentration per µL DM, non-DM, and PMI PRP (p= 0.22). However, non-DM PRP contained significantly lower VEGF per 1000 lysed platelets (0.20 (0.04−0.35) fg) compared to DM (0.69 (0.21−1.17) fg, p= 0.03) and PMI PRP (1.84 (1.38−2.10), p= 0.01), with no significant difference between DM and PMI PRP (p=0.06). The number of viable stromal cells per gram adipose tissue and collonies generated from DM SVF were significantly lower than non-DM (86.35 (52.48−106.76) x 10⁶ vs 158.93 (101.59−185.94) x 10⁶, p= 0.01 and 94 ± 14 collonies vs 31 ± 32 collonies, p= 0.004). Non-significant differences were also observed in the percentage of viable stromal cells (p= 0.24), expression of CD73 (p= 0.21), CD90 (p= 0.90), adipogenesis, chondrogenesis, osteogenesis, P2 and P3 PDT (p= 0.27 and 0.21, respectively), and the percentage of viable P2 ADSC (p= 0.07), but the expression of CD105 of P1 DM ADSC (64.41 (51.20−73.38)% vs 91.40 (82.62−95.47)%, p< 0.001) and the percentage of viable P3 ADSC (82.70 ± 8.07% vs 91.15 ± 3.77%, p= 0.04) were significantly lower than non-DM ADSC. The reduction of mRNA VEGF and VEGFR2 relative expression of P3 DM ADSC (0.64 (0.30−1.08) p= 0.86 and 0.64 ± 0.56, p= 0.49, respectively) were unsignificant compared to non-DM ADSC. Mean of VEGF level normalized to 1x10³ viable cells in the conditioned medium (CM) of DM and non-DM ADSC were 0.74 pg/mL and 0.62 pg/mL, respectively. Optimum 15% PRP-preconditioned DM ADSC had significantly lower PDT value (2.33 ± 0.56 days vs 5.04 ± 1.26 days, p=0.01) and higher percentage of viable cells compared to control (95.53 ± 1.60% vs 78.95 ± 10.13%, p=0.01). The increase of relative expression of CD105, mRNA VEGF and VEGFR2 (1.81 ± 0.73, p= 0.01; 5.27 ± 5.69, p= 0.23; and 9.01 ± 11.59, p= 0.06, respectively) were unsignificant in DM ADSC compared to non-DM. Optimum 15% PRP-preconditioned DM ADSC and control secreted VEGF in CM as much as 0.57 pg/mL and 1.67 pg/mL per 1x10³ viable cells in average. PRP-preconditioning improved the capillary tube formation in DM ADSC, but the process was longer compared to control, and unsignificant to non-DM ADSC and PRP-preconditioned DM ADSC with anti-VEGF (p=0.78).

Conclusions: Cellular damage in DM ADSC was idientified by a reduction of proliferation, differentiation. mRNA VEGF and VEGFR2 expression, and angiogenic potency. Preconditioning DM ADSC with 15% PRP (VEGF 98.00 pg/mL) improved the cellular damage with synergitic effect of VEGF and other growth factors contained in PRP.

 

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"Latar belakang. Latihan fisik yang dijalankan secara teratur dengan intensitas dan durasi tertentu akan merangsang remodeling jantung sebagai usaha untuk mempertahankan fungsi ventrikel terhadap peningkatan beban biomekanik pada jantung. Diperkirakan bahwa latihan fisik jangka panjang menimbulkan remodeling jantung menyerupai hipertrofi kardiomiopati, berupa hipertrofi miosit dengan kekacauan tatanan miosit, fibrosis, apoptosis dan ko-lokalisasi gap junction. Tujuan penelitian. Mengetahui dampak latihan fisik jangka panjang dan henti-latih pada remodeling kardiomiosit. Metode penelitian. Penelitian eksperimental in vivo pada tikus Wistar ini dilakukan di Departemen Biokimia dan Biologi Molekuler, Laboratorium Imunohistologi Departemen Patologi Anatomi dan Bagian Fisiologi, FKUI. Latihan fisik dengan intensitas dan jangka waktu latihan yang berbeda, serta henti latih setelah periode latihan diterapkan pada tikus Wistar jantan dewasa muda. Dilakukan analisis terhadap perubahan morfologi kardiomiosit, fibrosis, apoptosis (ekspresi Caspase-3, Bax dan Bcl-2), gap junction (ekspresi Connexin43) dan pola EKG. Perubahan morfologi kardiomiosit diamati menggunakan pulasan Hematoxylin Eosin, fibrosis diamati menggunakan pulasan Masson?s Trichrome, sedangkan ekspresi Caspase-3, Bax, Bcl-2 dan Connexin43 diamati melalui pulasan imunohistokimia. Rekaman EKG dilakukan dengan filter 100 Hz, pada kecepatan kertas 50 mm/detik dan kepekaan 1 mV = 20 mm. Hasil dan pembahasan. Hasil penelitian ini menunjukkan bahwa latihan aerobik dan anaerobik menimbulkan hipertrofi eksentrik dengan peningkatan fibrosis dan apoptosis serta ko-lokalisasi gap junction ke arah lateral membran. Perubahan kardiomiosit, peningkatan fibrosis dan apoptosis lebih nyata pada latihan anaerobik dibandingkan latihan aerobik. Pola EKG menunjang adanya pembesaran ventrikel akibat latihan aerobik dan anaerobik disertai gangguan repolarisasi yang nyata terutama pada latihan anaerobik. Henti latih tidak mengembalikan morfologi miosit, apoptosis dan lokalisasi gap junction ke keadaan semula. Pola EKG setelah periode henti latih pada latihan aerobik tetap menunjukkan adanya hipertrofi ventrikel tanpa gangguan penghataran impuls yang berarti, namun pada latihan anaerobik tetap didapatkan gangguan repolarisasi berupa pemanjangan interval QTc yang bermakna. Kesimpulan. Remodeling kardiomiosit akibat latihan fisik jangka panjang tidak menyerupai struktur hipertrofi kardiomiopati, namun disertai peningkatan apoptosis, kolokalisasi Cx43 dan gangguan penghantaran impuls. Henti-latih tidak memulihkan remodeling jantung maupun gangguan penghantaran impuls listrik.

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Background. Regular physical exercise with certain intensity and duration stimulates remodeling of the heart as an effort to preserve ventricular function against an increased biomechanical load. It is postulated that long-term exercise induces cardiac remodeling that resemble cardiomyopathy hypertrophy marked by cardiomyocyte disarray, fibrosis, apoptosis and co-localization of gap junction. Research objective. To study the effect of long-term physical exercise and detraining on cardiomyocyte remodeling.

Methodology. This in vivo experimental study was conducted at the Departement of Biochemistry and Molecular Biology, Immunohistology Laboratory of Departement of Pathological Anatomy and Departement of Physiology FMUI. Physical exercise with different intensity and periods of training was performed in groups of young adult male Wistar rats, followed by a period of detraining. Analysis of cardiomyocyte morphological changes, fibrosis, apoptosis (expression of Caspase-3, Bax and Bcl-2), gap junctions (expression Connexin43) and ECG pattern was conducted. Changes in cardiomyocyte morphology was observed using Haematoxylin Eosin staining, fibrosis was observed using Masson's Trichrome staining, whereas the expression of Caspase-3, Bax, Bcl-2 and Connexin43 was observed through immunohistochemical staining. ECG recording was done with a filter of 100 Hz, the paper speed of 50 mm/sec and the sensitivity of 1 mV = 20 mm.

Results and discussion. The results showed that aerobic and anaerobic exercises cause the development of eccentric hypertrophy, with increased fibrosis and apoptosis as well as co-localization of gap junction to the lateral site of the membrane. Cardiomyocyte remodeling, fibrosis and apoptosis were more prominent in anaerobic compared to aerobic exercise group. The ECG pattern supports enlargement of the ventricles due to aerobic and anaerobic exercises with noticeable repolarization disturbances, especially in the anaerobic group. Detraining did not return myocyte morphology, apoptosis and localization of gap junctions to its basic state. The ECG pattern after a period of detraining following aerobic exercise supports the existence of ventricular hypertrophy without significant disturbances in impulse conduction, however repolarization disturbances in the form of significant prolongation of QTc interval persist in the group with anaerobic exercise.

Conclusion. Cardiomyocyte remodeling due to long-term physical exercise did not resemble cardiomyopathy hypertrophy structure, although increased apoptosis, colocalization of Cx43 and distrbances in impuls conduction were observed. Detraining did not restore cardiac remodeling and disturbances in electrical impulse conduction."

"ABSTRAKLatar Belakang: Trombositosis pada pasien kanker payudara KPD diduga berkontribusi pada penyebaran dan sifat invasi sel punca kanker payudara. Modifikasi lingkungan mikro tumor dapat dilakukan untuk meningkatkan efektivitas terapi anti kanker. Belum diketahui apakah platelet derived growth factor PDGF -AB dalam lisat trombosit LT juga berperan terhadap cancer stem cell CSC payudara CD24-/CD44 .Tujuan: Penelitian ini bertujuan untuk menganalisis efek LT dan PDGF-AB didalamnya sebagai lingkungan mikro tumor pada proliferasi dan sifat invasi sel punca kanker payudara CD24-/CD44 yang ditandai dengan kadar matrix metalloproteinase-9 MMP-9 dan epithelial-cadherin E-cadherin . Metode: Penelitian ini merupakan studi eksperimental pada kultur sel punca KPD yang diberi LT dari pasien KPD dan donor sehat. Darah semua donor dilakukan pemeriksaan hematologi dan diproses untuk mendapatkan platelet rich plasma PRP . Jumlah trombosit per ?L PRP setiap donor dihitung. PRP diproses untuk mendapatkan LT. Kadar PDGF-AB LT diukur. LT 0,01 ditambahkan ke dalam medium dulbecco rsquo;s modified eagle rsquo;s medium DMEM -F12 untuk kultur sel punca KPD. Setelah inkubasi 48 jam, total jumlah sel, population doubling time PDT dan viabilitas sel dihitung dan dinormalisasikan terhadap nilai kontrolnya. Ekspresi MMP-9 dan E-cadherin dipilih sebagai penanda biologi sifat invasi dan diukur dengan metode enzyme-linked immunosorbent assay ELISA . Jumlah total sel, PDT, viabilitas sel, kadar MMP-9 dan E-cadherin dibandingkan antara pasien KPD dan donor sehat lalu dianalisis korelasinya dengan jumlah trombosit dan kadar PDGF-AB dalam lisat trombosit. Hasil: Jumlah trombosit dan kadar PDGF-AB dalam LT pasien KPD lebih tinggi dibandingkan LT donor sehat, keduanya dengan nilai p=0,02. LT pasien KPD memicu proliferasi sel punca KPD lebih baik dibandingkan LT donor sehat p

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ABSTRACTBackground Thrombocytosis in breast cancer BC patient is supposed to play a role in the invasiveness of breast cancer stem cells. Modification of tumor microenvironment was proposed to increase the efficacy of anticancer therapy. Aim This study aimed to analyze the effect of platelet lysate PL as well as its platelet derived growth factor PDGF AB content as a tumor microenvironment on the CD24 CD44 breast cancer stem cell BCSC proliferation and invasiveness. Methods This experimental study treated BCSC culture with PL from BC patients or healthy donors. Venous blood from all donors were subjected to hematology test and processed to obtain PRP. Platelet counts in PRP were determined. PRP was processed to obtain PL. PDGF AB contents in PL were measured. PL 0.01 was supplemented into dulbecco rsquo s modified eagle rsquo s medium DMEM F12 medium and used for culturing the CD24 CD44 BCSCs . After 48 hours, total cell count, population doubling time PDT , and cell viability were calculated and normalized to its control. Matrix metalloproteinase 9 MMP 9 and E cadherin was used as biological marker for CSC invasiveness and measured by enzyme linked immunosorbent assay ELISA method. Total cell count, PDT, cells viability as well as MMP 9 and E cadherin levels between BCSC, healthy donor platelet lysate and control group were compared and their correlation with platelet count in PRP and PDGF AB levels in platelet lysates were analyzed. Results Platelet counts and PDGF AB levels were higher in BC patient PL compared to healthy donor group, both with a p value of 0.02. BC patient PL could stimulate the proliferation of BCSCs higher than healthy donor PL p"

"Latar belakang: Cedera saraf perifer merupakan komplikasi trauma ekstremitas pada 3-10 pasien yang menyebabkan kelainan fungsi normal saraf. Terapi sel punca mesenkimal MSC telah banyak dikembangkan untuk regenerasi sel dan jaringan. Conditioned medium CM yang berasal dari tali pusat MSC-TP manusia dalam regenerasi saraf perifer belum banyak diketahui. Penelitian ini bertujuan untuk membandingkan perbaikan fungsi dan struktur saraf perifer.Metode: Penelitian ini merupakan penelitian eksperimental menggunakan 36 ekor tikus putih Rattus norvegicus berumur 2-3 bulan dengan berat badan berkisar 250-300 g. Penelitian dilakukan di laboratorium RSCM-FKUI dan Pusat Penelitian dan Pengembangan selama 3 tahun. Hewan coba dibagi menjadi 3 kelompok, yaitu kelompok kontrol atau sham SH , terapi standar ST , dan perlakuan conditioned medium MSC-TP CM . Penelitian dibagi menjadi dua jangka waktu, yaitu jangka pendek 7 hari pasca operasi HPO dan jangka panjang 70 HPO . Pemeriksaan yang dilakukan adalah analisis berjalan, elektrofisiologi dan imunohistokimia.Hasil: Hasil pemeriksaan fungsi motorik SFI, TFI, PFI, Q1-Q4 dan TOA , kelompok CM menunjukkan kesembuhan yang lebih cepat dibanding kelompok ST. Hasil gambaran elektrofisiologi, kelompok CM memiliki kecepatan konduksi saraf NCV lebih baik dibandingkan kelompok ST. Berdasarkan gambaran histologis, pewarnaan HE menunjukkan jumlah sel saraf yang lebih banyak pada kelompok CM dibanding kelompok ST pada hari ke-7 HPO dan 70 HPO. Pewarnaan TB menunjukkan diameter selubung mielin yang lebih tebal pada kelompok CM dibandingkan kelompok ST pada hari ke-7 HPO dan 70 HPO. Marker CD34 menunjukkan jumlah pembuluh darah memiliki hasil pada kelompok CM yang mendekati kelompok SH pada hari ke-7 HPO dan 70 HPO. Marker S100 menunjukkan persentase yang lebih banyak pada kelompok CM dibanding kelompok ST pada hari ke-7 HPO dan 70 HPO.Kesimpulan: CM MSC-TP mampu memberikan pengaruh terhadap perbaikan struktur dan fungsi saraf perifer pascacedera saraf pada kelompok CM hari ke-7 HPO.

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Background Peripheral nerve injury is a complication of extremities trauma in 3 10 of patients causing the dysfunction of nerve. Mesenchymal stem cell conditioned medium MSC CM is used as therapy to regenerate cells and tissues. However, the ability of human umbilical cord derived mesenchymal stem cell conditioned medium HU MSC CM in regenerating peripheral nerves has not been known. This research aimed to compare the functional and structural repairs of peripheral nerve.

Method This study is an experimental research using 36 rats of Sprague Dawley Rattus norvegicus strain aged 2 3 months with body weight ranging from 250 to 300 grams. The research was conducted in various laboratories at RSCM FKUI and the Center for Health Research and Development for three years. The experimental animals were divided into 3 groups, namely the control group SH , the standard therapy treatment group ST , and the conditioned medium treatment group CM . The research was divided into two stages consisting of a short term research of 7 days of post surgery PS and a long term research 70 PS . The examinations performed were in the form of motor function for the walking analysis, electrophysiology, and immunohistochemistry.

Result Based on the examination of motor function SFI, TFI, PFI, Q1 Q4, and TOA , the CM group showed faster recovery compared to the ST group. Based on the electrophysiological images, the CM group was able to have a better nerve conduction velocity NCV compared to the ST group. Based on the histological images, HE staining showed a higher amount of all nerve cells in the CM group compared to the amount in the ST group on the 7th day of PS and 70th day of PS. TB staining showed a thicker myelin sheath diameter in the CM group than that in the ST group on the 7th day of PS and 70th day of PS. CD34 marker showed that the number of blood vessels of the CM group was close to that of the SH group on the 7th day of PS and 70th day of PS. S100 marker had a higher percentage in the CM group compared to the ST group on the 7th day of PS and 70th day of PS.

Conclusion HU MSC CM is able to affect the functional and structural repairs of the peripheral nerve after nerve injury in the CM group on the 7th day of PS."

"Latar Belakang: Sel punca mesenkimal SPM asal jaringan lemak ASCs dan tali pusat UCSCs merupakan sumber sel punca yang umum digunakan pada terapi seluler. SPM berkomunikasi dengan sel kanker diantaranya melalui berbagai faktor biologis aktif yang dinamakan sekretom. Lingkungan mikro kanker yang hipoksik dapat memberi pengaruh berbeda pada interaksi ini. Efek interaksi sekretom SPM terhadap agresivitas sel punca kanker payudara hingga kini belum banyak diketahui. Tujuan: Menganalisis berbagai penanda agresivitas yang berkaitan dengan pertumbuhan dan ketahanan hidup viabilitas, proliferasi, sifat pluripotensi OCT4 dan SOX2, tumorigenik MFU, progresif-agresif TGF-?1 dan T?R1, penanda kepuncaan dan kemampuan detoksifikasi ALDH1A1 dan ALDH1A3, serta sifat invasif MMP2 dari sel punca kanker payudara BCSCs ALDH paska interaksi dengan sekretom dari conditioned medium CM SPM asal tali pusat dan jaringan adiposa yang diproduksi dalam kondisi normoksia maupun hipoksia. Metode: Studi eksperimental in vitro menggunakan CM UCSCs dan ASCs normoksia dan hipoksia yang disuplementasikan pada medium asal DMEM-F12 dari sel punca kanker payudara BCSCs ALDH dengan konsentrasi 50 v/v selama 72 jam. Analisis uji viabilitas dan proliferasi, q-RT-PCR ekspresi mRNA ALDH1A1, ALDH1A3, OCT4, SOX2, MMP2, TGF-?1 dan T?R1 serta uji MFU dari BCSCs ALDH dilakukan untuk mengetahui efek dari sekretom dalam CM terhadap agresivitas BCSCs ALDH. Hasil: Sekretom dalam CM-UCSCs dapat meningkatkan agresivitas BCSCs melalui peningkatan penanda invasif ndash;agresif dan detoksifikasi. Sekretom dalam CM-ASCs dapat meningkatkan agresivitas BCSCs melalui peningkatan penanda pluripotensi, invasif dan detoksifikasi. Prekondisi hipoksia pada CM-SPM dapat meningkatkan potensi agresivitas lebih tinggi daripada CM normoksia. Perbedaan regulasi viabilitas dan proliferasi serta turunnya penanda tumorigenik BCSCs paska suplementasi CM perlu diinterpretasikan dengan hati-hati dan masih memerlukan verifikasi. Kesimpulan: Sekretom dalam CM UCSCs maupun ASCs dapat memberikan efek meningkatkan sifat agresif dari BCSCs ALDH. Preparasi hipoksia pada produksi CM cendrung lebih mendukung sifat agresif dari BCSCs ALDH dibandingkan CM normoksianya. Perbedaan regulasi viabilitas dan proliferasi serta turunnya penanda tumorigenik pada BCSCs paska suplementasi CM SPM masih membutuhkan penelitian lebih lanjut untuk menganalisis dasar molekuler yang menyebabkannya.

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Background: Adipose and umbilical cord tissue derived mesenchymal stem cells MSCs are the most common sources that are used in various cellular therapies. MSCs are known to communicate with cancer cells through their secretomes. The hypoxic microenvironment of cancer may cause different effects on this interaction. Effects of MSC secretomes against the aggressiveness of breast cancer stem cells BCSCs ALDH have not been widely investigated.

Aim: To analyze various markers of aggressiveness that are associated with growth and survival viability, proliferation, pluripotency OCT4 and SOX2, tumorigenic MFU, progressive aggressive TGF 1 and T R1, stemness and detoxification ALDH1A1 and ALDH1A3, as well as the invasive nature MMP2 of BCSCs ALDH post interaction with both normoxic and hypoxic MSC secretomes.

Methods: The in vitro experimental study using conditioned medium CM of MSCs produced in normoxic and hypoxic condition that were supplemented in medium of BCSCs ALDH with concentrations of 50 v v for 72 hours. Analysis of viability, proliferation, and q RT PCR of ALDH1A1, ALDH1A3, OCT4, SOX2, MMP2, TGF 1 and T R1 mRNA as well as MFU assay were performed to determine the effect of secretomes on the aggressiveness of BCSCs ALDH.

Results: Secretomes of UCSCs supported the aggressiveness by increasing invasive aggressive and detoxification markers, while secretomes of ASCs supported the aggressiveness by increased pluripotency, invasive and detoxification markers. Hypoxic preconditioning of MSC secretomes increased the potential for aggressiveness higher than normoxic secretomes. Differences in viability and proliferation regulation and the decrease in BCSCs tumorigenic post secretomes supplementation need to be interpreted carefully and further verification.

Conclusion: Supplementation of MSC secretomes increased the aggressive properties of BCSCs ALDH. The hypoxic secretomes tend to favor the aggressive nature of BCSCs ALDH compared to its counterpart. Differences in viability and proliferation regulation as well as the decrease in tumorigenic markers in BCSCs after MSC secretomes supplementation still need further research to analyze the molecular underlying basis."

"kegagalan hati. Organoid hati dapat digunakan sebagai bahan untuk BAL. Organoid hati merupakan rekonstruksi hati kultur 3D dari sel punca dan sel-sel lainnya yang menyerupai mikrostruktur hati in vivo dan memiliki fungsi hati. Organoid hati juga dapat digunakan untuk uji obat dan sebagai model unutk mempelajari penyakit hati.Metode : Organoid hati pada penelitian ini direkonstruksi dari hepatosit, sel stelata hepatika (LX2), sel punca mesenkimal asal tali pusat (UC-MSCs), dan sel punca hematopoiesis asal darah tali pusat (UCB-CD34+). Hepatosit primer tikus, LX2, UC-MSCs dan UCB-CD34+ diko-kultur dalam 11 formulasi rasio selama 2 hari. Formulasi rasio yang membentuk sferoid dikultur dalam 4 medium kultur selama 5 hari, dipanen dan dilakukan analisa viabilitas. Rasio dengan viabilitas tertinggi merupakan rasio optimal dalam medium kultur optimal untuk rekonstruksi organoid hati. Rasio hepatosit : LX2 : UC-MSCs : UCB-CD34+ optimal 5 : 1 : 2 : 2 diko-kultur dalam medium kultur optimal Williams E yang disuplementasi dengan PRP, ITS dan dexamethasone selama 14 hari dan dilakukan analisa morfologi, fungsi hati dan potensi angiogenesis.Hasil : Viabilitas organoid bertahan hingga hari ke-14 dan ekspresi protein albumin, ekspresi protein GOT dan ekspresi protein CD31 cukup stabil hingga hari ke-14. Ekspresi gen Albumin meningkat hingga hari ke-14 sedangkan ekspresi gen GOT menurun hingga hari ke-14. Sekresi urea menurun hingga hari ke-5 dan sekresi albumin menurun hingga hari ke-7.Kesimpulan : Organoid hati ini direkonstruksi dari hepatosit primer, LX2, UC-MSCs, UCB-CD34+ dengan rasio optimal 5 : 1 : 2 : 2 dalam medium kultur optimal sederhana dan ekonomis Williams E yang disuplementasi PRP, ITS dan dexamethasone. Organoid hati ini dapat mempertahankan viabilitas dan fungsi hingga hari ke-14. Organoid hati penelitian ini dapat digunakan sebagai model untuk uji obat dan dapat dikembangkan untuk menjadi bahan BAL.

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Introduction : Bioartificial Liver (BAL) is being developed to be an alternative therapy for liver failure. Liver organoids can be used as prototype material for BAL. Liver organoids are 3D cultured liver reconstructions of stem cells and other cells that resemble the liver microstructure in vivo and perform liver function. Liver organoids also can be used for drug testing and as a model for liver disease pathogenesis.

Methods : Liver organoids in this study were reconstructed from hepatocytes, hepatic stellate cells (LX2), human umbilical cord-mesenchymal stem cells (UC-MSCs), and human umbilical cord blood (UCB) hematopoiesis stem cells CD34+. Rat primary hepatocytes, LX2, UC-MSCs and UCB-CD34+ were co-cultured in 11 ratio formulations for two days. The ratio formed spheroid were cultured in four culture medium for five days, harvested and analyzed for viability. The ratio with the highest viability was the optimal ratio in the optimal culture medium for hepatic organoid reconstruction. The optimal ratio 5 : 1: 2 : 2 of Hepatocytes : LX2 : UC-MSCs : UCB-CD34+ was co-cultured in optimal culture medium Williams E supplemented with PRP, ITS and dexamethasone for 14 days and analyzed for morphology, liver function and angiogenesis potential.

Results : Liver organoids viability maintained until day 14 and albumin protein expression, GOT protein expression and CD31 protein expression were quite stable until day 14. Albumin gene expression increased until day 14, while GOT gene expression decreased until day 14. Urea secretion decreased until day 5 and albumin secretion decreased until day 7. Conclusion : These liver organoids were reconstructed from optimal ratio 5 : 1 : 2 : 2 of primary hepatocytes, LX2, UC-MSCs, UCB-CD34+ in simple and economical optimal culture medium Williams E supplemented by PRP, ITS and dexamethasone. These liver organoids maintained viability and liver function until day 14. These liver organoids can be used as a model for drug testing and can be developed to become a BAL material for future application."

"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."