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"Latar Belakang: Meningkatnya angka kejadian stroke dan beratnya disabilitas dari penderita stroke yang bertahan hidup, menjadikan diperlukannya terapi yang optimal untuk restorasi paska stroke. Neurorestorasi dengan transplantasi sel punca menjanjikan perbaikan luaran fungsional yang baik pada pasien stroke iskemik. Penelitian ini dibuat untuk mengamati efek transplantasi Stem Cell from Human Exfoliated Deciduous Teeth SHED pada luaran klinis dan rasio sel neuron mati pada model stroke iskemik, untuk mendapatkan terapi yang optimal untuk stroke iskemik.Metode: Pembuatan tikus model stroke iskemik dilakukan dengan oklusi arteri cerebri media (MCAO). Pada 48 jam setelah MCAO, dilakukan transplantasi sel mesenkimal asal SHED secara intravena, dengan dosis 2x106/kgBB. Dilakukan evaluasi fungsional tikus secara neurobehaviour dengan tes Y Maze, dan evaluasi sensorimotor tikus dengan tes silinder. Evaluasi rasio neuron mati dilakukan dengan pewarnaan Hematoksilin dan Eosin.Hasil: Terdapat perbaikan evaluasi neurobehaviour dengan Y Maze (p=0,04) dan evaluasi sensorimotor dengan tes silinder (p=0,04) pada 14 hari setelah transplantasi pada kelompok tikus yang ditransplantasi SHED dibandingkan kontrol. Terdapat pengurangan rasio sel neuron mati (p=0,0) pada tikus yang ditransplantasi SHED pada 21 hari setelah MCAO.Kesimpulan: Transplantasi SHED pada tikus model stroke iskemik pada fase akut stroke menunjukkan perbaikan klinis dan terdapat pengurangan rasio neuron mati pada otak tikus model stroke iskemik yang di transplantasi dengan sel mesenkimal asal SHED.

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Background: The incidence of stroke reaches 15 million cases worldwide, and 5 million stroke survivors suffered permanent disability. Ischemic stroke causes a burden on health problems particularly in Indonesia. The prevalence of stroke in Indonesia in 2013 is 7 per 1000 population. The optimal stroke restoration therapy required, and neurorestoration with stem cell transplantation is a promising therapy that provides functional improvements for ischemic stroke. This research was conducted to observe the effects of Stem Cell from Human Exfoliated Deciduous Teeth (SHED) transplantation on the clinical improvement and neuron death ratio in the brain of rats models with ischemic stroke.Methods: One group of normal rats and two groups (n=5) of male wistar rats undergone permenents Middle Cerebral Artery Occlusion (MCAO). SHED transplantation performed 48 hours after MCAO, by intravenous injection with a dose of 2x106 cells/kg. Functional evaluation conducted in rats with Y Maze and cylinder Test. Evaluation of the death neurons ratio in brain cortex area done by Hematoksilin and Eosin staining.Results : The functional evaluation using Y Maze and Cylinder Test was significantly improved in the treatment group compared to the control stroke group p < 0,05 14 days after MCAO. There was a reduction in the neuron death ratio p = 0.0 in rats transplanted with SHED.Conclusion : SHED transplantation in acute stroke showed clinical improvement and reduction in the neuron death ratio in the brain of rat models with ischemic stroke. Keywords: Cell transplantation, Ischemic Stroke, MCAO, SHED"

"Latar belakang : Anemia akut sering terjadi pada anak sakit kritis yang dirawat diPICU, memiliki konsekuensi hipoksia global yang dapat mengakibatkan disfungsimiokardium. Transfusi PRC masih menjadi salah satu pilihan dalam rangkamemperbaiki oksigenasi dan kinerja jantung saat terjadi anemia. Bukti-bukti pengaruhtransfusi pada perbaikan performa jantung masih terbatas.Tujuan : Mengevaluasi kadar NT-proBNP, pasokan oksigen, indeks inotropi dan rasioenergi potensial:energi gerak pada jantung sebelum dan sesudah transfusi PRC padaanak sakit kritis yang mengalami anemia akut.Metode : Penelitian analitik observasional potong lintang sejak April sampai Agustus2019 pada anak usia 1 bulan-18 tahun dengan sakit kritis yang dirawat di PICURSUPN Dr. Cipto Mangunkusumo. Penilaian hemodinamik menggunakan USCOM.Hasil : Penelitian ini melibatkan 31 subyek dengan median umur 3,6 tahun (rentang0,1-17,5 tahun). Kadar Hb naik sebesar 29,1±15,9% setelah mendapat transfusi PRC9±3,3 mL/KgBB. Rerata kadar hemoglobin sebelum dan sesudah transfusi adalah7,94±1,46 dan 10,17±1,92 g/dL (p<0,000 IK 95%: 1,80-2,64). Kadar NT-proBNPmeningkat tak bermakna sebesar 12% (-77,0-199) setelah transfusi dari 4214±6678menjadi 5182±8327 pg/mL (p=0,186 IK 95%: -493-2428). Tidak terdapat korelasiantara persen perubahan Hb dan NT-proBNP (Spearman correlation r=0,124; p=0,505).Terdapat kenaikan pasokan oksigen pasca transfusi sebesar 20,7±38,9% dan berkorelasidengan kanaikan hemoglobin (Pearson correlation r=0,39; p=0,029). Uji Chi-squaremenunjukkan adanya hubungan bermakna antara kelompok yang mengalami kenaikanDO dengan perbaikan indeks inotropi (uji Chi square, p=0,031) dan perbaikan PKR(p=0,008), namun tak ada hubungan dengan perubahan NT-proBNP (p=0,511).Simpulan : Tidak terdapat perubahan bermakna kadar NT-proBNP sebelum dansesudah transfusi PRC pada anak sakit kritis yang mengalami anemia akut. Peningkatanpasokan oksigen pasca transfusi PRC berkorelasi dengan peningkatan indeks inotropi(Smith-Madigan Inotropy Index) dan perbaikan potensial to kinetic ratio (PKR)

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Background: Acute anemia often occurs in critically ill children in PICU, which hasglobal hypoxic consequences resulting myocardial dysfunction. Transfusion of PRC isstill choosen in order to improve oxygenation and cardiac performance during anemia.Evidence of the effect of transfusion on improving cardiac performance is still limited.Objective: To evaluate NT-proBNP levels, delivery oxygen (DO2), inotropy index andthe potential to kinetic energy ratio (PKR) of heart before and after PRC transfusion incritically ill children with acute anemia.Methods: A cross-sectional observational analytic study conducted from April toAugust 2019 in children aged 1 month-18 years cared in PICU Dr. CiptoMangunkusumo Hospital. Hemodynamic assessment using USCOM.Results: This study involved 31 subjects with a median age of 3.6 years (range 0.1-17.5years). Hb levels increased by 29.1±15.9% after receiving a 9±3.3 mL / KgBBtransfusion PRC. The mean hemoglobin levels before and after the transfusion were7.94±1.46 and 10.17±1.92 g / dL (p <0.000; CI 95%: 1.80-2.64). NT-proBNP levelsslight increased but not statistically sgnificant by 12%(-77.0 - 199) after PRCtransfusion from 4214±6678 to 5182±8327 pg/mL (p = 0.186; CI 95%: -493 - 2428).There was no correlation between percent change in Hb and NT-proBNP (Spearmancorrelation r=0.124; p=0.505). There was increasing in DO2 after transfusion by20.7±38.9% and correlated with increased hemogolobin (Pearson correlation r=0.39;p=0.029). Chi-square test showed a significant relationship between groups thatexperienced an increase in DO2 with an improvement in the inotropy index (Chi squaretest, p=0.031) and improvement in PKR (p=0.008), but there was no relationship withNT-proBNP changes (p=0.511) .Conclusions: There was no significant change in NT-proBNP levels before and afterPRC transfusion in critically ill children who had acute anemia. Increased DO2 afterPRC transfusion correlates with an increase in the inotropy index (Smith-MadiganInotropy Index) and improvement in potential to kinetic ratio (PKR)."

"This volume aims to cover important aspects of the various facets of organ transplantation and regenerative medicine."

"Advances in stem cell research discusses recent advances in stem cell science, including therapeutic applications. This volume covers such topics as biomanufacturing iPS cells for therapeutic applications, techniques for controlling stem cell fate decisions, as well as current basic research in such areas as germ line stem cells, genomics and proteomics in stem cell research. "

"ABSTRAKLatar belakang. Eksitoksisitas merupakan salah satu mekanisme penting dalam kerusakan otak masa perinatal. Eksitoksisitas terjadi akibat peningkatan glutamat ekstrasel. Stem cell From Human Exfoliated Deciduous dapat mensekresi enzim GAD yang akan memgkatalisis perubahan glutamat menjadi GABA. Perubahan glutamat menjadi GABA menyebabkan kadar glutamat ekstrasel menurun, namun peningkatan GABA dapat menyebabkan terjadinya depolarisasi yang dapat berakibat timbulnya eksitoksisitas. penelitian bertujuan untuk menentukan potensi neuroproteksi CM SHED mencegah kerusakan progenitor neuron akibat induksi glutamat.Metode. Progenitor neuron diisolasi dari otak tikus umur 2 hari dan conditioned medium didapat dari MSC SHED. Sampel dibagi menjadi 4 kelompok, yaitu kelompok kultur progenitor neuron dengan medium neurobasal tanpa glutamat dan glisin N- , dengan medium neurobasal ditambah glutamat dan glisin N , dengan CM SHED tanpa glutamat dan glisin K- , dengan CM SHED ditambah glutamat dan glisin K . Pada progenitor neuron dilakukan pemeriksaan kadar mRNA GABAAR1, subunit NR2B NMDAR dengan RT PCR, kadar protein NMDAR1 dan GABAAR1 dengan ELISA. Caspase -3 dan 7AAD dengan Muse. Pada CM dilakukan pemeriksaan kadar GABA dengan Elisa.Hasil. Viabilitas progenitor neuron pada kelompok K 78,05 lebih tinggi dari kelompok kontrol N- 73,22 , sedangkan kelompok N lebih kecil 68,90 . Kelompok K memiliki kadar GABA paling tinggi dan berbeda bermakna dengan kelompok lainnya, sedangkan pada kelompok N paling kecil. Kadar GABA yang tinggi pada kelompok K diduga karena adanya enzim GAD yang dapat mengkatalisis perubahan glutamat menjadi GABA pada CM SHED. Hasil pemeriksaan mRNA GABAAR1 kelompok K paling tinggi dibandingkan kelompok lain, diduga disebabkan GABA dapat mengaktivasi jalur MAPK yang menyebabkan terjadinya proses transkripsi mRNA GABAAR1. Kadar protein GABAAR1 pada kelompok K paling kecil dibandingkan kelompok lain, hal ini diduga disebabkan karena adanya perubahan subunit dari 2 akibat peningkatan kadar GABA. Kadar GABA yang rendah pada kelompok N diduga disebabkan GABA banyak terpakai terikat dengan reseptor GABAAR1, sehingga terlihat rendah saat pemeriksaan. Hasil ini sesuai dengan pemeriksaan kadar GABAAR1, dimana pada kelompok N kadar GABAAR1 paling tinggi. Kemungkinan lain adalah adanya umpan balik negatif oleh GABAAR1. Pada pemeriksaan mRNA GABAAR1 kelompok N didapat penurunan ekspresi relatif terhadap kontrol. Diduga peningkatan protein GABAAR1 sudah tidak diperlukan sehingga ekspresi mRNA GABAAR1 dihambat. Pemeriksaan dilakukan setelah perlakuan selama 24 jam, diduga kondisi progenitor neuron telah mengalami fase akhir. Terdapat perubahan dinamik pada regulasi GABAAR1 progenitor neuron. Pada fase awal aktivitas eksitatorik merupakan hasil kerjasama GABAAR1 dan NMDAR, kemudian terjadi perubahan aktivitas GABAAR1 dari eksitatorik menjadi inhibitorik. Pada kelompok N diduga aktivitas inhibitorik GABAAR1 tidak dapat mengatasi aktivitas eksitatorik akibat induksi glutamat, sehingga menyebabkan terjadinya apoptosis. Sedangkan pada kelompok K , dimana terdapat peningkatan GABA akibat CM SHED, aktivitas inhibitorik GABAAR1 dapat mengimbangi aktivitas eksitatorik oleh glutamat sehingga terjadi neuroproteksi.Kesimpulan. Dari hasil penelitian ini dapat disimpulkan CM SHED memiliki potensi neuroproteksi dalam mencegah apoptosis progenitor neuron akibat induksi glutamat.

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ABSTRACTBackgound. Exitoxicity is one of the important mechanisms in perinatal period brain damage. Exitoxicity results from increased extracellular glutamate. Stemcell From Human Exfoliated Deciduous can secrete a GAD enzyme that will analyze the change of glutamate to GABA. The change of glutamate to GABA causes extracellular glutamate levels to decrease, but an increase in GABA can lead to depolarization which may result in the excitoxicity. the study aimed to determine the potential neuroprotection of CM SHED to prevent progenitor neuron damage of glutamate induction.Method. Progenitor neurons were isolated from the brains of mice aged 2 days and conditioned medium obtained from MSC SHED. The samples were divided into 4 groups, ie the progenitor culture group of neurons with neutral glutamate and neurobasal medium N- , with neurobasal medium plus glutamate and glycine N , with CM SHED without glutamate and glycine K- , with CM SHED plus glutamate and glycine K . In progenitor neuron, GABAAR1 mRNA, NR2B NMDAR subunit with PCR RT, protein NMDAR1 and GABAAR1 with ELISA were examined. Caspase -3 and 7AAD with Muse. In CM, GABA levels were evaluated with Elisa.Result. The viability of progenitor neurons in the K group 78.05 was higher than the control group N 73.22 , whereas the N group was smaller 68.90 . The K group had the highest levels of GABA and was significantly different from the other groups, whereas in the smallest N group. High GABA levels in the K group are thought as a result of the presence of GAD enzymes that can catalyze the change of glutamate to GABA in CM SHED. The highest level of GABAAR1 group compared to other groups was thought to be caused by GABA to activate MAPK pathway causing transcription of GABAAR1 mRNA. The level of GABAAR1 protein in the K group was the smallest compared to the other groups, presumably due to a subunit change from ? 2 due to elevated levels of GABA. Low levels of GABA in the N group are thought to be caused by GABA being widely used bound with GABAAR1 receptors, making it noticeably low during examination. This result is in accordance with the GABAAR1 concentration, which in the N group of GABAAR1 levels is highest. Another possibility is negative feedback by GABAAR1. On examination of group GABAAR1 mRNA N , there was a decrease in expression relative to control. It is suspected that the increase in GABAAR1 protein is not needed so that GABAAR1 mRNA expression is inhibited. The examination was performed after 24 hours treatment, presumably progenitor neuron has ending phase. There is a dynamic change in GABAAR1 progenitor neuron regulation. In the early phase of excitatory activity is the result of cooperation GABAAR1 and NMDAR, then there is a change of activity GABAAR1 from excitatory become inhibitory. In the N group it is suspected that GABAAR1 inhibitory activity can not overcome the excitatory activity by caused of glutamate induction, thus causing apoptosis. While in the K group, where there is an increase in GABA considering CM SHED, GABAAR1 inhibitory activity can compensate for the excitatory activity by glutamate resulting in neuroprotection.Conclution. From the results of this study can be concluded CM SHED has the potential of neuroprotection in preventing progenitor neuron apoptosis through glutamate induction."

"Pertumbuhan kanker tidak hanya ditentukan oleh adanya sel kanker itu sendiri akan tetapi ditentukan juga oleh lingkungan mikro disekitarnya. Lingkungan mikro tersebut merupakan jaringan yang heterogen dengan adanya interaksi sel termasuk sel punca mesenkim. Sekretom mengandung faktor-faktor biologis terlarut yang dapat mempengaruhi pertumbuhan sel kanker. Stem cell from Human Exfoliated Deciduous SHED diketahui merupakan sumber sel punca yang memiliki banyak potensi. Sampai saat ini belum diketahui bagaimana dampak pemberian CM dari SHED terhadap sel punca kanker payudara. Oleh karena itu, penelitian ini bertujuan untuk menganalisis pemberian conditioned medium kultur SHED pada sel punca kanker payudara terhadap viablitas, proliferasi dan tumorigenitas serta kepuncaan dari sel punca kanker payudaraALDH dan sel punca kanker MCF7. Conditioned medium SHED SHED-CM adalah medium kultur bebas serum sel SHED yang dikumpulkan dalam 24 jam dan 48 jam. ALDH dan MCF7diberikan 50 v/v SHED-CM 24 jam dan 48 jam dan di inkubasi selama 72 jam. Hal yang sama dilakukan untukperlakuan aktivasi CM dengan suhu. CM sebelum digunakan terlebih dahulu dipanaskan dalam suhu 80 C selama 10 menit. Kontrol adalah sel yang diberikan 50 v/v a-MEM. Perhitungan viabilitas sel dilakukan dengan menggunakan metode Trypan Blue Exclusion Assay dan ekspresi relatif mRNA dari TGF-b1, TGF-b1 receptor TBRI, ALDH1A1 dan OCT4 menggunakan qRT-PCR dan analisis menggunakan perhitungan livak. Hasil penelitian menunjukkan bahwa ekspresi relatif dari TGF-b1, TGF-b1 reseptor TBRI, ALDH1A1 dan OCT4 pada sel ALDH dan MCF7 pasca induksi dengan CM SHED mengalami peningkatan yang signifikan dibandingkan dengan kontrol. Selain itu, peningkatan yang lebih signifikan ditunjukkan pada perlakuan aktivasi dibandingkan yang tidak diaktivasi. Hal yang berbeda pada hasil uji viabilitas sel. Viabilitas sel mengalami penurunan pasca induksi dengan CM SHED sedangkan setelah diinduksi dengan CM SHED yang telah diaktivasi, viabilitas sel mengalami peningkatan yang signifikan pada sel ALDH dan MCF7. Dengan demikian sekretom SHED dapat meningkatkan viabilitas dan proliferasi serta kemampuan kepuncaan dari ALDH dan MCF7.

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Cancer development is not only determined by corresponding of cancer cells but also by the microenvironment. This includes a heterogen network of interacting cell include mesenchymal stem cells. Conditioned medium of MSC culture containing soluble factor hes been identified to affect intercellular communicating between MSC and cancer cells which could affect the stemness of cancer cells. Many studies reported that Stem cells from human exfoliated deciduous teeth SHED as a novel stem cell source with multipotent potential. However, the effect of MSC interaction with cancer cells can not be clearly understood so that the effects of safety in its utilization are not yet known for certain. This study is to confirm the relation between secretom of MSC from SHED with the stemness and agresiveness of ALDH and MCF7. SHED conditioned medium SHED CM, SHED were grown in serum free a MEM for 24 and 48 hours, consist of two groups, non heated and heated at 80 C for 10 min. Human BCSCs ALDH cultured in DMEM F12 were supplemented with 50 v v CM SHED 24 h and 48 h, as well as with heated by 72 h incubation. Control was BCSCs supplemented with 50 v v a MEM. We measured the viability with trypan blue assay and mRNA expression of TGF b1, TGF b1 receptor TBRI, as well as stemness genes ALDH1A1 and OCT4 using qRT PCR. The relative mRNA expression levels of TGF b1, T RI, OCT4 and ALDH1A1 in BCSCs supplemented with non heated SHED CM were increased compared to their control and also after TGF b1 heat activation was significantly higher than in non heated SHED CM. In the other hand, the viability cell was significantly reduced after supplemented with non heated SHED CM, but increased higher than control when treated with heated SHED CM, there are may be a role of the TGF b1 signaling involvement of other factors in SHED CM affect cell proliferation and increase the stemness. We found that secretomes SHED can increase proliferation of breast cancer stem cells ALDH and also expression of stemness gene OCT4 and ALDH1A1. the activation with heated can enhance the increase of proliferation and stemness. We assumed that signalling of TGF can affect tumor proggresion of ALDH and MCF7"

"Human dental pulp of exfoliated deciduous teeth contains the population of cells that exhibited mesenchymal stem cell (MSC) characters. Though, a cell amplification process is indeed required to secure and adequate cell number for such a potential employment. Several publications suggested the alteration of MSCs upon in vitro culture, for example, the decrease in proliferation and the loss of stem cell characters. Here, we investigated an influence of basic fibroblast growth factor (bFGF) on stem cells isolated from human exfoliated deciduous teeth (SHEDs) with respect to cell proliferation, colony forming unit efficiency and stem cell marker expression in both short- and long-term cultures. For short-term bFGF treatment, SHEDs were treated with bFGF for 48h. While, in long-term bFGF supplementation, SHEDs were maintained in culture and continuous passage upon confluence in medium supplemented witg bFGF. Cells at passage (P) 5 and 10 were employed for characterization. Our result showed that short-term bFGF treatment enhanced OCT4, REX1, and NANOG mRNA expression as well as colony forming unit ability. The FGFR inhibitor pretreatment was able to attenuate the influence of bFGF on pluripotent stem cell marker expression, confirming bFGF function. In addition, cells cultured in high passage number had decreased in cell proliferation, colony forming unit capacity, and pluripotent stem cell marker mRNA expression. However, bFGF supplementation in culture medium enhanced both pluripotent stem cell marker expression and colony forming unit capacity in later passage, though the effect was not robust. Together, these results indicate that high passage number may attenuate pluripotent properties of SHEDs and bFGF supplementation could be the beneficial approach to maintain SHEDs' stemness properties. "

"ABSTRACTPenelitian ini menganalisis kemampuan Human Serum Albumin (HSA), Umbilical cord blood (UCB) serum dan Fetal Bovine Serum (FBS) dalam menjaga stabilitas ekspansi ex vivo kultur sel punca hematopoetik (SPH). Sel yang digunakan adalah sel mononuklear dan sel CD34+ dari darah tali pusat yang disimpan beku dalam lingkungan nitrogen. Medium basal kultur yang digunakan adalah RPMI 1640 Biowest dan Stemspan. Jumlah sel hidup dihitung menggunakan metode eksklusi tryphan blue dan fenotipe sel CD34+ dianalisis menggunakan flow cytometry. Pewarnaan Giemsa dilakukan pada sel-sel yang dipanen pada hari ketujuh kultur untuk menganalisis morfologi sel. Besar sampel dalam penelitian ini adalah tiga dan jumlah pengulangan adalah dua kali. Penelitian ini menunjukkan bahwa kultur dengan suplementasi HSA menghasilkan jumlah sel yang lebih rendah namun memiliki persentase CD34+ yang lebih tinggi dibandingkan UCB serum dan FBS. Pewarnaan Giemsa menunjukkan sel-sel darah yang terdiferensiasi paling sedikit ditemukan pada HSA. Hasil tersebut menunjukkan bahwa, medium dengan suplementasi HSA lebih unggul dari UCB serum dan FBS dalam mempertahankan kepuncaan sel punca hematopoetik.

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ABSTRACTThis study analyzed the ability of serum Human Serum Albumin (HSA), Umbilical cord blood (UCB) and Fetal Bovine Serum (FBS) to maintain the stability of ex vivo expansion of hematopoietic stem cell (SPH) cultures. The cells used are mononuclear cells and CD34 + cells from cord blood which are frozen in a nitrogen environment. The basal culture medium used was RPMI 1640 Biowest and Stemspan. The number of living cells was calculated using the tryphan blue exclusion method and the CD34 + cell phenotype was analyzed using flow cytometry. Giemsa staining was carried out on cells harvested on the seventh day of culture to analyze cell morphology. The sample size in this study was three and the number of repetitions was twice. This study shows that culture with HSA supplementation results in lower cell counts but has a higher CD34 + percentage compared to serum UCB and FBS. Giemsa staining shows the least differentiated blood cells are found in HSA. These results indicate that, medium with HSA supplementation is superior to serum UCB and FBS in maintaining hematopoietic stem cell stem cells."