Isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan in vivo = Isolation culture and characterization of human osteosarcoma in vitro and in vivo study
| Main Author: | Waluyo Sugito, author |
|---|---|
| Format: | Bachelors Doctoral |
| Terbitan: |
, 2016
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| Subjects: | |
| Online Access: |
http://lib.ui.ac.id/file?file=digital/2017-4/20435264-SP-Waluyo Sugito.pdf |
| ctrlnum |
20435264 |
|---|---|
| fullrecord |
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<dc schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd"><title>Isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan in vivo = Isolation culture and characterization of human osteosarcoma in vitro and in vivo study</title><creator>Waluyo Sugito, author</creator><type>Thesis:Bachelors</type><place/><publisher/><date>2016</date><description><b>ABSTRAK</b><br>
Pendahuluan. Pemahaman dan publikasi mengenai aspek biologi sel osteosarkoma manusia di Indonesia masih terbatas, sehingga dibutuhkan suatu penelitian tentang isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan pada model hewan secara in vivo. Penelitian ini bertujuan untuk mengetahui apakah sel osteosarkoma manusia dapat diisolasi dan dikultur secara in vitro dan secara in vivo pada hewan model tikus Sprague Dawley (SD).
Metode Penelitian. Pada tahap pertama dilakukan isolasi dan kultur sel osteosarkoma dari 6 pasien pre-kemoterapi neoadjuvant dan 4 pasien telah mendapat kemoterapi neoadjuvant. Isolasi dan kultur dengan metode eksplant. Karakterisasi sel osteosarkoma dibuktikan dengan pemeriksaan morfologi sel, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) dan imunositokimia. Tahap kedua menghasilkan model hewan tikus SD dengan inokulasi sel hasil kultur 1.106 sel per ekor ke intramedular femur distal (3 ekor) dan ke intramuskular gastroknemius dan soleus tibia proksimal (3 ekor). Pengukuran Alkali fosfatase serum dilakukan pada minggu ke-0, 4, dan 8, radiologi pada minggu ke-4 dan ke-8 dan histopatologi dilakukan pada minggu ke-8.
Temuan Penelitian. Sitologi menunjukkan sel tumor dengan inti yang pleiomorfik, hiperkromatik, letak di tepi dan anak inti nyata. Pemeriksaan RT-PCR menunjukkan ekspresi gen positif terhadap marker STAT3, Nanog, OCT3/4 dan CD 133. Pada pemeriksaan IFA didapatkan hasil positif terhadap antibodi osteokalsin, alkali fosfatase, dan CD 133. Pada pemeriksaan imunositokimia didapatkan hasil positif terhadap antibodi alkali fosfatase dan osteokalsin. Tahap kedua, evaluasi radiologi tidak menunjukkan gambaran destruksi tulang maupun tumbuhnya massa pada soft tissue. Pada histopatologi gambaran jaringan yang normal.
Simpulan. Sel osteosarkoma dapat diisolasi dan dikultur dari jaringan tumor pasien osteosarkoma serta menunjukkan karakterisasi sel sesuai gambaran osteosarkoma pada pasien penderita osteosarkoma. Belum dapat dilakukan pembuatan hewan model osteosarkoma dari hewan Tikus Sprague Dawley immunocompetent.
<hr>
<b>ABSTRACT</b><br>
Introduction. Understanding and publications about biological aspect of human osteosarcoma cells in Indonesia are scarce, so study about isolation, culture and characterization of by in vitro or in vivo are needed. This study aimed to understand whether human osteosarcoma cells could be isolated and cultured by in vitro and in vivo for animal model Sprague Dawley (SD) rat.
Methods. First stage, isolation and culture of osteosarcoma cell from 6 patients with neoadjuvant prechemotherapy and 4 that already received neoadjuvant chemotherapy. Isolation and culture used explant method. Characterization used morphological examination of cells, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) and immunocytochemistry. Objective of second stage was producing animal models experiment by inoculation of 1.106 cells intra medullary distal femur (3 animals) and to intramuscular gastrocnemius and soleus proximal tibia (3 animals). Serum alkaline phosphatase was checked at week 0,4 and 8, radiology week 4 and 8, and histopathology at week 8.
Results. Cytology showed tumor cell with pleiomorphic, hyperchromatic nucleus on the edge and conspicuous nucleoli. RT-PCR examination was positive for gene expression in STAT3 marker, Nanog, OCT 3/4 and CD 133. IFA was positive for osteocalcin, alkaline phosphatase, and CD 133 antibodies. In immunocytochemical examination there were positive result of alkaline phosphatase and osteocalcin antibody. For second stage, radiology evaluation showed no bone destruction or mass growth in soft tissue. Histopathology showed normal tissue.
Conclusions. Osteosarcoma cell could be isolated and cultured from osteosarcoma?s patient and showed cell characterization that corresponded with the picture of osteosarcoma cell in patient with osteosarcoma. Animal model of osteosarcoma in immunocompetent SD rat could not be done.</description><subject>Osteosarcoma -- Treatment</subject><identifier>20435264</identifier><source>http://lib.ui.ac.id/file?file=digital/2017-4/20435264-SP-Waluyo Sugito.pdf</source><recordID>20435264</recordID></dc>
|
| format |
Thesis:Bachelors Thesis Thesis:Doctoral |
| author |
Waluyo Sugito, author |
| title |
Isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan in vivo = Isolation culture and characterization of human osteosarcoma in vitro and in vivo study |
| publishDate |
2016 |
| topic |
Osteosarcoma -- Treatment |
| url |
http://lib.ui.ac.id/file?file=digital/2017-4/20435264-SP-Waluyo Sugito.pdf |
| contents |
<b>ABSTRAK</b><br>
Pendahuluan. Pemahaman dan publikasi mengenai aspek biologi sel osteosarkoma manusia di Indonesia masih terbatas, sehingga dibutuhkan suatu penelitian tentang isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan pada model hewan secara in vivo. Penelitian ini bertujuan untuk mengetahui apakah sel osteosarkoma manusia dapat diisolasi dan dikultur secara in vitro dan secara in vivo pada hewan model tikus Sprague Dawley (SD).
Metode Penelitian. Pada tahap pertama dilakukan isolasi dan kultur sel osteosarkoma dari 6 pasien pre-kemoterapi neoadjuvant dan 4 pasien telah mendapat kemoterapi neoadjuvant. Isolasi dan kultur dengan metode eksplant. Karakterisasi sel osteosarkoma dibuktikan dengan pemeriksaan morfologi sel, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) dan imunositokimia. Tahap kedua menghasilkan model hewan tikus SD dengan inokulasi sel hasil kultur 1.106 sel per ekor ke intramedular femur distal (3 ekor) dan ke intramuskular gastroknemius dan soleus tibia proksimal (3 ekor). Pengukuran Alkali fosfatase serum dilakukan pada minggu ke-0, 4, dan 8, radiologi pada minggu ke-4 dan ke-8 dan histopatologi dilakukan pada minggu ke-8.
Temuan Penelitian. Sitologi menunjukkan sel tumor dengan inti yang pleiomorfik, hiperkromatik, letak di tepi dan anak inti nyata. Pemeriksaan RT-PCR menunjukkan ekspresi gen positif terhadap marker STAT3, Nanog, OCT3/4 dan CD 133. Pada pemeriksaan IFA didapatkan hasil positif terhadap antibodi osteokalsin, alkali fosfatase, dan CD 133. Pada pemeriksaan imunositokimia didapatkan hasil positif terhadap antibodi alkali fosfatase dan osteokalsin. Tahap kedua, evaluasi radiologi tidak menunjukkan gambaran destruksi tulang maupun tumbuhnya massa pada soft tissue. Pada histopatologi gambaran jaringan yang normal.
Simpulan. Sel osteosarkoma dapat diisolasi dan dikultur dari jaringan tumor pasien osteosarkoma serta menunjukkan karakterisasi sel sesuai gambaran osteosarkoma pada pasien penderita osteosarkoma. Belum dapat dilakukan pembuatan hewan model osteosarkoma dari hewan Tikus Sprague Dawley immunocompetent.
<hr>
<b>ABSTRACT</b><br>
Introduction. Understanding and publications about biological aspect of human osteosarcoma cells in Indonesia are scarce, so study about isolation, culture and characterization of by in vitro or in vivo are needed. This study aimed to understand whether human osteosarcoma cells could be isolated and cultured by in vitro and in vivo for animal model Sprague Dawley (SD) rat.
Methods. First stage, isolation and culture of osteosarcoma cell from 6 patients with neoadjuvant prechemotherapy and 4 that already received neoadjuvant chemotherapy. Isolation and culture used explant method. Characterization used morphological examination of cells, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) and immunocytochemistry. Objective of second stage was producing animal models experiment by inoculation of 1.106 cells intra medullary distal femur (3 animals) and to intramuscular gastrocnemius and soleus proximal tibia (3 animals). Serum alkaline phosphatase was checked at week 0,4 and 8, radiology week 4 and 8, and histopathology at week 8.
Results. Cytology showed tumor cell with pleiomorphic, hyperchromatic nucleus on the edge and conspicuous nucleoli. RT-PCR examination was positive for gene expression in STAT3 marker, Nanog, OCT 3/4 and CD 133. IFA was positive for osteocalcin, alkaline phosphatase, and CD 133 antibodies. In immunocytochemical examination there were positive result of alkaline phosphatase and osteocalcin antibody. For second stage, radiology evaluation showed no bone destruction or mass growth in soft tissue. Histopathology showed normal tissue.
Conclusions. Osteosarcoma cell could be isolated and cultured from osteosarcoma?s patient and showed cell characterization that corresponded with the picture of osteosarcoma cell in patient with osteosarcoma. Animal model of osteosarcoma in immunocompetent SD rat could not be done. |
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