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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊台鴻(Tai-Horng Young) | |
| dc.contributor.author | Yi-Chen Li | en |
| dc.contributor.author | 李亦宸 | zh_TW |
| dc.date.accessioned | 2021-06-16T16:27:57Z | - |
| dc.date.available | 2018-12-31 | |
| dc.date.copyright | 2013-02-01 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-01-11 | |
| dc.identifier.citation | [1] McKay R. Stem cells in the central nervous system. Science 1997;276:66-71.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63201 | - |
| dc.description.abstract | 神經幹/前驅細胞可以在懸浮於基材表面上的情況下增生並形成神經幹/前驅細胞球，它被認為是一種非下錨型的細胞。然而，神經幹/前驅細胞球則被認為是在適當條件下貼附於基材上時，會進而分化成不同種類細胞的一種下錨型細胞。本研究的目的是藉由結合培養基和生醫材料來調控神經幹/前驅細胞的增生與分化。眾所皆知血清中具有許多促進增生或分化的分子可以對神經幹/前驅細胞的行為產生極大的影響。因此本論文中分為兩的主要的部份。 第一部份為探討血清的角色，10%的胎牛血清及其部份成份被加至培養基之去評估對於胚胎鼠大腦皮質所分離的神經幹/前驅細胞的分化潛能之影響。當神經幹/前驅細胞被分化到七天之後，利用免疫螢光染色去標定神經系細胞的特定蛋白，可以發現到當培養基中含有血清中小於分子量100 kDa的分子和存在纖維母細胞生長因子時可以促進神經幹/前驅細胞往神經元的分化。相反的，神經幹/前驅細胞則是在存在全血清成份時，其分化的細胞主要為膠質細胞。由半定量來分析免疫染色的結果，當神經幹/前驅細胞存在於結合血清成份小於100 kDa、聚乙烯乙烯醇共聚物 (EVAL)和纖維連接蛋白可以發現爬出球外的細胞具有神經元表現的比例被促進到約有85%之多，此外藉由中腦中央動脈阻塞的動物中風實驗模型， 也可以發現到分清的成份，分子量小100 kDa 可以促進神經元的生成及中風動物的行為功能回復。 論文中的第二部份是10%的胎牛血清及其成份被加至培養基之去評估對於胚胎鼠大腦皮質所分離的神經幹/前驅細胞的貼附及增生潛能之影響。文中顯示上皮生長因子和纖維連接蛋白對神經幹/前驅細胞具有一個協同效應。而當血清成份存在於培養基中時，結合上皮生長因子和纖維連接蛋白比其單一成份對神經幹/前驅細胞的影響有明顯的差別。有趣地當血清中存在上皮生長因子和纖維連接蛋白時，神經幹/前驅細胞會快速地貼附於細胞培養盤上而且爬出來的細胞有高於70%維持著巢蛋白的表現。而這些在具有貼附形態的未分化巢蛋白表現的神經幹/前驅細胞仍可以被誘導分化成其他神經系的細胞。這些結果提供了結合了上皮生長因子和纖維連接蛋白可以有效地將神經幹/前驅細胞在貼附的情況下維持巢蛋白的表現，和可以做為一個線索去研究血清成份對神經幹/前驅細胞不分化的反應。更進一步地，結合上皮生長因子、纖維連接蛋白和高分子導管，也可以讓神經幹/前驅細胞在導管中維持不分化的狀態，這樣顯示可能可以被使用在神經導管的應用上。這些結果是令人鼓舞的， 因為當想要一個神經元分化或是維持神經幹細胞不分化的環境時，我們可以在神經科學研究的領域中提供有用的策略去控制神經發育的過程。 | zh_TW |
| dc.description.abstract | Neural stem/precursor cells (NSPCs) can proliferate in suspension to form neurospheres that do not need to attach to the substrate surface and, thus, can be considered as anchorage-independent cells. However, neurospheres are considered to be anchorage-dependent cells, which differentiate into different cell types when they attach to the substrate under appropriate conditions. The purpose of this study was to regulate the proliferation and differentiation of NSPCs by the combination of media, biomaterials. It is known that many proliferation- and differentiation-promoting molecules are present in the serum, which has a great effect on the behaviors of NSPCs. There are two major parts in this thesis. One is that considering the role of serum, 10% fetal bovine serum or its fractions were added to DMEM/F12 medium to examine the effect of the differentiation-promoting potential on cultured NSPC isolated from embryonic rat cerebral cortex. The NSPCs were cultured for 7 days, after which differentiation was assayed using immunocytochemistry for neural lineage specific markers. It was demonstrated that molecules promoting neuron differentiation were present in serum with molecular weight <100 kDa, which could dominate the differentiation of NSPCs principally into neurons in the presence of basic fibroblast growth factor (bFGF). In contrast, NSPCs were induced to differentiate predominantly into glial cell phenotypes in the presence of whole serum components. Based on medium containing serum fraction, semi-quantification showed that the MAP2-positive percentage of the immunoreactive ratio within migrated cells could be promoted over 85% by combining poly(ethylene-co-vinyl alcohol) (EVAL) biomaterial and fibronectin matrix protein. In addition, by the middle central artery occlusion (MCAO) rat, It also be demonstrated that the component of serum with molecular weight <100 kDa could promote the generation of neuron in the ischemia brain and recover the function of MCAO rat.
The other part in the thesis is that 10% FBS or its components were added into culture medium to examine the effect of the adhesion- and proliferation-promoting potential on NSPCs. It appeared that medium containing epidermal growth factor (EGF) and fibronectin had a synergistic effect to regulate NSPCs, and cooperation of EGF and fibronectin was more significant than singly effect in regulation of NSPCs when serum components were present in the culture system. Interestingly, when culture medium was in the presence of EGF and fibronectin, NSPCs rapidly attached onto the TCPS surface and more than 70% migrating-out cells were predominantly maintaining nestin-positive phenotype. Furthermore, the result of undifferentiated nestin-positive dominating the pattern of NSPCs adhesion could be induced to differentiate into neural linage cells. These results informed that the combined administration of serum components with EGF and fibronectin could significantly maintain nestin-positive of adhesive NSPCs phenotype, and will therefore be an important clue to investigate what molecules in serum are responsible for the NSPCs undifferentiation-maintaining activity. Furthermore, combination of the medium containing EGF and fibronectin and polymer conduit also could maintain NSPCs at undifferentiated phenotype in the conduit, which might be applied to be a neuronal conduit. These results are very encouraging, since an environment favorable for neuronal differentiation and maintenance of undifferentiated NCPSs should be useful in the development of strategies for controlling the behavior of NSPCs in neuroscience research. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T16:27:57Z (GMT). No. of bitstreams: 1 ntu-102-F95548001-1.pdf: 28469760 bytes, checksum: c123aa84ccd9a303029c7c069a24b939 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | List of Contents
致謝……………………………………………………… I 摘要……………………………………………………………………………IV Abstract………………………………………………………VI List of Contents………………………………………………………IX List of Figures………………………………………………………XII Abbreviations………………………………………………………XVII Chapter 1 Introduction………………………………………………………1 1.1 Background and Objectives………………………………………………………1 1.2 Stem cells…………………………………………………………………………2 1.3 Central nervous systems…………………………………………………………2 1.4 NSPCs from CNS…………………………………………………………………4 1.5 Cell-soluble factors interactions………………………………………………7 1.6 Cell-substrate interaction………………………………………………………8 1.7 Polymer membranes………………………………………………………………9 1.7.1 PVDF………………………………………………………11 1.7.2 EVAL………………………………………………………11 1.7.3 PVA………………………………………………………12 1.7.4 Chitosan………………………………………………………12 1.7.5 PCL………………………………………………………13 1.8 Application of NSPCs………………………………………………………14 Chapter 2 Materials and Methods………………………………………………………15 2.1 Flowchart of experiment design…………………………………………………15 2.2 Materials…………………………………………………………………………16 2.3 Instruments………………………………………………………………………19 2.4 Solution preparation……………………………………………………………20 2.4.1 Phosphate buffered saline (PBS)…………………………………………………20 2.4.2 Hank’s balanced salt solution (HBSS)………………………………………………………20 2.4.3 4% paraformaldehyde/0.5% Triton X-100………………………………………………………21 2.4.4 Trypsin………………………………………………………22 2.4.5 Western Blot………………………………………………………22 2.5 Preparation of membranes………………………………………………………24 2.6 Preparation of polymer conduit………………………………………………24 2.7 Prepare ECM-coated substrates…………………………………………………26 2.8 Isolation and culture of NSPCs………………………………………………26 2.9 Preparation of serum fraction…………………………………………………27 2.10 Immunocytochemistry…………………………………………………………28 2.11 Flow cytometry…………………………………………………………………29 2.12 Western blot analysis……………………………………………………………29 2.13 Labeling and semi-quantitative analysis of functionally active synapses………30 2.14 Alamar blue assay………………………………………………………………31 2.15 Scratch assay……………………………………………………………………31 2.16 Animal middle cerebral artery occlusion model and injection…………………31 2.17 TTC staining……………………………………………………………………32 2.18 Behaviors testing………………………………………………………………32 2.18.1 Rotarod testing………………………………………………………32 2.18.2 Neurological Severity Score testing………………………………………………………33 2.18.3 Body asymmetry testing………………………………………………………33 2.19 Scanning electron microscope………………………………………………………33 Chapter 3 Results………………………………………………………34 3.1 Part I Fundamental behaviors of NSPCs………………………………………………………34 3.1.1 Cell characterization………………………………………………………34 3.1.2 Cell differentiation………………………………………………………34 3.2 Part II NSPCs with different medium components………………………………37 3.2.1 Effect of different medium components on NSPCs………………………………………………………37 3.2.2 Effect of combination of different medium components or growth factors on NSPCs………………………………………………………37 3.2.3 Behavior of neurospheres cultured in the presence of serum fraction with molecular weights <100 kDa………………………………………………………40 3.2.4 Synapyic functions of differentiated neurons migrated out from neuropsheres in the presence of serum fraction with molecular weights <100 kDa………………………………………………………47 3.2.5 Behavior of neurospheres cultured in the presence of 100KD medium and whole serum………………………………………………………50 3.3 Part III A new method for NSPCs culture………………………………………53 3.3.1 Behavior of neurospheres cultured in the presence of serum components with molecular weights > 100 kDa………………………………………………………53 3.3.2 Behavior of neurospheres cultured in the medium with fibronectin and different growth factors………………………………………………………54 3.3.3 Behavior of neurospheres cultured in the medium with fibronectin and EGF………………………………………………………60 3.3.4 The effect of medium B on NSPC proliferation………………………………………………………67 3.3.5 The effect on the proliferation and differentiation capabilities of medium B-treated NSPC after removing EGF and fibronectin………………………………………………………71 3.3.6 The effect on the wound healing capabilities of medium B-treated NSPC………………………………………………………75 3.3.7 The effect on neurospheres in medium B with different concentrations of EGF and fibronectin………………………………………………………79 3.3.8 The adhesion force of neurospheres with the treatment of EGF and fibronectin…………………………………………………………………84 3.3.9 The mechanism of neurospheres with the treatment of EGF and fibronectin………………………………………………………89 3.4 Part IV Application………………………………………………………93 3.4.1 A in vitro neuronal differentiation system………………………………………………………93 3.4.2 Stroke recovery………………………………………………………113 3.4.3 A in vitro neuronal conduit system………………………………………………………131 Chapter 4 Discussion………………………………………………………140 4.1 100KD medium and its application………………………………………………………140 4.2 Medium B and its application………………………………………………………148 Chapter 5 Conclusions………………………………………………………159 References………………………………………………………161 Supplemental Figures………………………………………………………180 Introduction of author………………………………………………………184 Education………………………………………………………185 Awards………………………………………………………186 Honors………………………………………………………186 Publications………………………………………………………187 Patents………………………………………………………190 專刊………………………………………………………191 Appendix………………………………………………………192 List of Figures Figure 1.4 1. The plasticity of NSPCs.………………………………………………………5 Figure 2.1 1. The flowchart of experiment design.………………………………………………………15 Figure 2.6 1. The process of preparing polymer conduit.………………………………………………………25 Figure 2.8 1. Isolation of NSPCs from E14-15 rat embryonic cerebral cortices. (a) The pregnant female wistar rat. (b) To cut the underbelly. (c) To isolate embryonic sacs from wistar rat. (d) To retrieve and immerse the embryos in HBSS solution. (e) To isolate the cortex and remove regions close to the mesenphalon.………………………………………………………27 Figure 2.18 1. Schematic representation of the time course of all functional recover design.………………………………………………………32 Figure 3.1 1. (a) Optical, (b) merged fluorescent, and (c, d) merged confocal photomicrographs of neurospheres cultured on TCPS in DMEM/F12 medium. Anti-nestin (green) and anti-Ki67 (red) were immunoreactive for undifferentiated NSPCs and proliferated cells, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………35 Figure 3.1 2. (a) Optical and (b, c, d) merged fluorescent photomicrographs of neurospheres cultured on TCPS in DMEM/F12 medium containing 10% FBS. Anti-GFAP (green), anti-MAP2 (red), and anti-O4 (yellow) was immunoreactive for differentiated astrocytes, neurons, and oligodendrocytes, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………36 Figure 3.2 1. Optical photomicrographs of neurospheres cultured on in DMEM/F12 medium with the treatment of albumin (5 mg/ml), fibronectin (1 μg/ml), insulin ( 10 ng/ml), urea (30 μg/ml), pretrescine (1 μM), or calcium (0.7 mM) after 7 days of incubation.………………………………………………………38 Figure 3.2 2. Optical photomicrographs of neurospheres cultured on in DMEM/F12 medium with the treatment of multicomponents (albumin (5 mg/ml), fibronectin (1 μg/ml), insulin ( 10 ng/ml), urea (30 μg/ml), pretrescine (1 μM), and calcium (0.7 mM)), growth factors (EGF (10ng/ml), bFGF (10ng/ml), IL-1α (10ng/ml), IL-1β (10ng/ml), IL-6 (10ng/ml), IGF-I (10ng/ml), IGF-2 (10ng/ml), PDGFAA (10ng/ml), PDGFAB (10ng/ml), PDGFBB (10ng/ml)), or combination of multicomponents and growth factors after 7 days of incubation.………………………………………………………39 Figure 3.2 3. Optical photomicrographs of neurospheres cultured on TCPS in DMEM/F12 medium containing 10% serum fraction with molecular weights <100 kDa (100KD), 50–100 kDa (50K–100K), and between <50 kDa (50KD) in the presence of bFGF after 7 days of incubation. Scale bar = 100 μm.………………………………………………………43 Figure 3.2 4. Fluorescent images of FM1-43-stained neurospheres, cultured on TCPS in 50 KD, 50 K–100 K and 100KD medium after 7 days of incubation, with puncta-like labeling of functional presynaptic terminals before and 150 s after the start of stimulation. For comparison, DMEM/F12 medium containing 10% whole serum (FBS) was included in this experiment. Scale bar = 100 μm.………………………………………………………48 Figure 3.2 5. Merged fluorescent images of neurospheres cultured on TCPS for 7 days as a function of the volume ratio of 100KD medium relative to whole serum (10%). Anti-MAP2 (red) and anti-GFAP (green) are immunoreactive for differentiated neurons and astrocytes, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………51 Figure 3.3 1. Optical photomicrographs of neurospheres cultured on TCPS in the medium F-M, or F/G after 7 day of incubation. F: fibronectin, G: globulin, M: α2-microglobulin, F/G: α2-microglobulin, fibronectin, and globulin. Scale bar = 100 μm.………………………………………………………53 Figure 3.3 2. Optical photomicrographs of neurospheres cultured on TCPS in the medium with fibronectin (1 μg/ml) and single growth factor (10 ng/ml) after 7 day of incubation. Scale bar = 100 μm.………………………………………………………55 Figure 3.3 3. Merged fluorescent images of neurospheres cultured on TCPS in the medium with fibronectin (1 μg/ml) and single growth factor (10 ng/ml) after 7 day of incubation. Scale bar = 100 μm.………………………………………………………56 Figure 3.3 4. Optical photomicrographs of neurospheres cultured on TCPS in the medium FBS, B, E, F, and DMEM/F12 after 7 day of incubation allow different behaviors of neurospheres. B: EGF/fibronectin, E: EGF, F: fibronectin. Scale bar = 100 μm.………………………………………………………62 Figure 3.3 5. Merged fluorescent images of neurospheres cultured on TCPS in the FBS after 7 day of incubation allow cell attachment and differentiation. Anti-nestin, anti-MAP2 and anti-GFAP are immunoreactive for undifferentiated neural stem/precursor cells, differentiated neurons and astrocytes, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………63 Figure 3.3 6. Optical photomicrographs of neurospheres cultured on TCPS in the FBS and medium B in 11 days of incubation. Scale bar = 100 μm.………………………………………………………68 Figure 3.3 7. Optical photomicrographs and (b) merged fluorescent images of medium B-treated cells reseeded new well with fresh serum-free DMEM/F12 medium after 6 days of incubation allow neurospheres reaggregating and maintaining NSPC phenotype. Anti-nestin (Green) is immunoreactive for undifferentiated neural stem/precursor cells, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………73 Figure 3.3 8. Optical photomicrographs and merged fluorescent images of neurospheres cultured on TCPS after changing from medium B to differentiation medium for 10 days of incubation allow NSPCs having various differentiation phenotypes. Anti-MAP2 (Red) and anti-GFAP (green) are immunoreactive for differentiated neurons and astrocytes, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………74 Figure 3.3 9. Optical photomicrographs of in vitro scratch assay of new neurospheres cultured on the scratched well after changing from medium B to differentiation medium (FBS or 100KD) in 24 days of incubation allow NSPCs having various differentiation abilities. Scale bar = 100 μm.………………………………………………………76 Figure 3.3 10. Optical photomicrographs of neurospheres cultured on TCPS in the different concentration of medium B after 7 day of incubation. 0.5Xmedium B: 5ng/ml EGF, 0.5 μg/ml fibronectin, 0.25Xmedium B: 2.5ng/ml EGF, 0.25 μg/ml fibronectin, 0.1Xmedium B: 1ng/ml EGF, 0.1 μg/ml fibronectin.………………………………………………………81 Figure 3.3 11. Optical photomicrographs of neurospheres after detachment for 5 min. neurospheres were pre-cultured on TCPS in the medium B, E, or F at 3 day of incubation. Scale bar = 100 μm.………………………………………………………85 Figure 3.3-13. Optical photomicrographs of neurospheres cultured on TCPS in the medium B in the presence or absence of U0126, a Erk inhibitor, after 7 day of incubation. Scale bar = 100 μm.………………………………………………………90 Figure 3.4 1. Optical photomicrographs of neurospheres cultured on PVDF, EVAL, PVA and TCPS in DMEM/F12 medium after 7 days of incubation. Scale bar = 100 μm.………………………………………………………94 Figure 3.4 2. Optical photomicrographs of neurospheres cultured on PVDF, EVAL, PVA and TCPS in DMEM/F12 medium containing 10% FBS after 7 days of incubation. Scale bar = 100 μm.………………………………………………………98 Figure 3.4 3. Optical photomicrographs of neurospheres cultured on PVDF, EVAL, PVA and TCPS in 100KD medium after 7 days of incubation. Scale bar = 100 μm.………………………………………………………102 Figure 3.4 4. Optical photomicrographs of neurospheres cultured on laminin-coated PVDF, EVAL, PVA and TCPS in 100KD medium after 7 days of incubation. Scale bar = 100 μm.………………………………………………………107 Figure 3.4 5. Optical photomicrographs of neurospheres cultured on fibronectin-coated PVDF, EVAL, PVA and TCPS in 100KD medium after 7 days of incubation. Scale bar = 100 μm.………………………………………………………110 Figure 3.4 6. The infarct sizes of a normal or MCAO brain of rat were examined by TTC staining.………………………………………………………115 Figure 3.4 7. Merged fluorescent images of cells at the infract region in the right brain of MCAO rat without the treatment of 100KD medium (control). Anti-MAP2 (Red) and anti-GFAP (green) are immunoreactive for differentiated neurons and astrocytes, respectively. DAPI (blue) was used to mark nuclei.………………………………………………………116 Figure 3.4 8. Merged fluorescent images of cells at the infract region in the right brain of MCAO rat without (control) or with the treatment of 100KD medium. Anti-nestin (green) was immunoreactive for undifferentiated NSPCs. DAPI (blue) was used to mark nuclei.………………………………………………………123 Figure 3.4 9. Merged fluorescent images of cell proliferation at the infract region in the right brain of MCAO rat without (control) or with the treatment of 100KD medium. Anti-Ki67 (green) was immunoreactive for proliferating cells. DAPI (blue) was used to mark nuclei.………………………………………………………126 Figure 3.4 10. The functional recovery in MCAO rats with or without the treatment of 100KD medium was estimated by (a) rotarod maintenance, (b) neurological deficit score, (c) body tilting percentage, and (d) body weight loss.………………………………………………………130 Figure 3.4 11. Optical photomicrographs of neurospheres cultured on EVAL and chitosan in the medium B after 7 day of incubation. Scale bar = 100 μm.………………………………………………………133 Figure 3.4 12. (a) PCL conduit and scanning electron micrographs of (b) external surface, (c) internal surface, and (d) cross-section of PCL conduit. Scale bar = 50 μm.………………………………………………………136 Figure 3.4 13. Scanning electron micrographs of neurospheres cultured in medium B in the PCL conduit after 7 days of incubation. (a) Low-power microscope field, scale bar = 500 μm; and (b) High-power microscope field, scale bar = 50 μm. (c) Low-power microscope field and (d) high-power microscope field merged fluorescent images of neurospheres cultured in medium B in the PCL conduit after 7 day of incubation. Anti-nestin, anti-MAP2 and anti-GFAP are immunoreactive for undifferentiated neural stem/precursor cells, differentiated neurons and astrocytes, respectively. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………139 Figure 4.1 1. The effects of 100 KD medium for NSPCs in vitro.………………………………………………………146 Figure 4.2 1. The mechanism of medium B for NSPCs.………………………………………………………158 Supplemental Figure 1.5 1. The components of fetal bovine serum.………………………………………………………180 Supplemental Figure 3.2-1. Optical photomicrographs of neurospheres cultured in 100 kDa serum fraction with different concentration of bFGF after 7 days of culture.………………………………………………………181 Supplemental Figure3.2-2. Neurospheres cultured on TCPS in DMEM/F12 medium in the presence of 20 ng/ml bFGF after 7 days of culture.………………………………………………………182 Supplemental Figure 4.2-1. Merged fluorescent images of cells at the infract region in the right brain of MCAO rat with the treatment of 100KD medium. Anti-β1-integrin (Red) was immunoreactive for β1-integrin. DAPI (blue) was used to mark nuclei. Scale bar = 100 μm.………………………………………………………183 | |
| dc.language.iso | en | |
| dc.subject | 神經幹/前驅細胞 | zh_TW |
| dc.subject | 上皮生長因子 | zh_TW |
| dc.subject | 纖維連接蛋白 | zh_TW |
| dc.subject | 血清 | zh_TW |
| dc.subject | 血清成份 | zh_TW |
| dc.subject | 未分化 | zh_TW |
| dc.subject | 分化 | zh_TW |
| dc.subject | 貼附 | zh_TW |
| dc.subject | 增生 | zh_TW |
| dc.subject | Neural stem/precursor cells | en |
| dc.subject | proliferation | en |
| dc.subject | adhesion | en |
| dc.subject | differentiation | en |
| dc.subject | undifferentiation | en |
| dc.subject | serum fraction | en |
| dc.subject | serum | en |
| dc.subject | fibronectin | en |
| dc.subject | EGF | en |
| dc.title | 生醫材料與培養基成份對神經幹細胞/前驅細胞行為之探討 | zh_TW |
| dc.title | Study on the effects of biomaterials and medium components on the behaviors of neural stem/precursor cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 賴君義(Juin-Yih Lai),王盈錦(Yng-Jiin Wang),鄭廖平(Liao-Ping Cheng),謝松蒼(Sung-Tsang Hsieh),劉得任(Der-Zen Liu) | |
| dc.subject.keyword | 神經幹/前驅細胞,上皮生長因子,纖維連接蛋白,血清,血清成份,未分化,分化,貼附,增生, | zh_TW |
| dc.subject.keyword | Neural stem/precursor cells,EGF,fibronectin,serum,serum fraction,undifferentiation,differentiation,adhesion,proliferation, | en |
| dc.relation.page | 196 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-01-11 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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