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- 技术资料
- 库存：
大量
- 年限：
1 day old newborn
- 物种来源：
其他
- 是否是肿瘤细胞：
0
- 细胞类型：
成纤维细胞
- 生长状态：
贴壁生长
- 器官来源：
肾脏
- 运输方式：
冻存运输
- 细胞形态：
成纤维样
- 相关疾病：
正常
- ATCC Number：
CCL-10™
| Designations: | BHK-21 [C-13] | ||
| Depositors: | I Macpherson | ||
| Biosafety Level: | 1 | ||
| Shipped: | frozen | ||
| Medium & Serum: | See Propagation | ||
| Growth Properties: | adherent | ||
| Organism: | Mesocricetus auratus | ||
| Morphology: | fibroblast |
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| Source: | Organ: kidney Disease: normal Cell Type: fibroblast |
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| Permits/Forms: | In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. | ||
| Isolation: | Isolation date: March, 1961 | ||
| Applications: | testing transfection host |
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| Virus Susceptibility: | Human adenovirus 25 Reovirus 3 Vesicular stomatitis virus Human poliovirus 2 |
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| Cytogenetic Analysis: | Chromosome Frequency Distribution 50 Cells: 2n = 44. This is a pseudodiploid line with the tetraploidy occurring at 4%. The karyotype is 44,XY,-6,-15,6q+,15q+ in a majority of cells analyzed. The markers 6q+ and 15q+ occurred in most cells. An occasional monosomic or trisomic condition for a normal chromosome was also detected. | ||
| Age: | 1 day old newborn | ||
| HeLa Markers: | N | ||
| Comments: | The parent line of BHK-21(C-13) was derived from the kidneys of five unsexed, 1-day-old hamsters in March, 1961, by I.A. Macpherson and M.G.P. Stoker. Following 84 days of continuous cultivation, interrupted only by an 8-day preservation by freezing, clone 13 was initiated by single-cell isolation. This line has been used as a host for transformation with expression vectors containing selectable and amplifiable marker DNAs (e.g., Factor VIII, see ATCC CRL-8544 ). The World Organization for Animal Health (OIE) uses the cells for routine diagnosis of rabies. (see: http://www.oie.int/Eng/Normes/Mmanual/A_00044.htm). |
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| Propagation: | ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C |
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| Subculturing: | Protocol: Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 is recommended Medium Renewal: 1 to 2 times per week |
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| Preservation: | Freeze medium: Complete growth medium 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase |
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| Related Products: | Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003 recommended serum:ATCC 30-2020 |
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| Bioreactive Factors: | Growth Factors: T cell growth factor (TCGF) | ||
| References: | 2: Drayna D, et al. Genetic mapping and diagnosis of haemophilia A achieved through a BclI polymorphism in the factor VIII gene. Nature 314: 738-740, 1985. PubMed: 2986011 3: Kazazian HH Jr., et al. Restriction site polymorphism in the phosphoglycerate kinase gene on the X chromosome. Hum. Genet. 66: 217-219, 1984. PubMed: 6325324 21411: Macpherson I, Stoker M. Polyoma transformation of hamster cell clones--an investigation of genetic factors affecting cell competence. Virology 16: 147-151, 1962. PubMed: 14468055 21412: . . Virology 14: 359-370, 1961. 25963: Macpherson I. Characteristics of a hamster cell clone transformed by polyoma virus. J. Natl. Cancer Inst. 30: 795-815, 1963. 27297: Deleersnyder V, et al. Formation of native hepatitis C virus glycoprotein complexes. J. Virol. 71: 697-704, 1997. PubMed: 8985401 32269: Yang TT, et al. Quantification of gene expression with a secreted alkaline phosphatase reporter system. BioTechniques 23: 1110-1114, 1997. PubMed: 9421645 32473: Hussain MA, et al. POU domain transcription factor brain 4 confers pancreatic alpha-cell-specific expression of the proglucagon gene through interaction with a novel proximal promoter G1 element. Mol. Cell. Biol. 17: 7186-7194, 1997. PubMed: 9372951 32475: You M, et al. ch-IAp1, a member of the inhibitor-of-apoptosis protein family, is a mediator of the antiapoptotic activity of the v-Rel oncoprotein. Mol. Cell. Biol. 17: 7328-7341, 1997. PubMed: 9372964 32501: Jelachich ML, Lipton HL. Theiler's murine encephalomyelitis virus kills restrictive but not permissive cells by apoptosis. J. Virol. 70: 6856-6861, 1996. PubMed: 8794327 32512: Schnell MJ, et al. The minimal conserved transcription stop-start signal promotes stable expression of a foreign gene in vesicular stomatitis virus. J. Virol. 70: 2318-2323, 1996. PubMed: 8642658 32524: Chang YE, et al. Properties of the protein encoded by the UL32 open reading frame of herpes simplex virus 1. J. Virol. 70: 3938-3946, 1996. PubMed: 8648731 92346: Biological evaluation of medical devices. Part 5: Tests for in vitro cytotoxicity. Sydney, NSW, Australia:Standards Australia;Standards Australia AS ISO 10993.5-2002. 92389: Biological evaluation of medical devices--Part 5: Tests for in vitro cytotoxicity. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 10993-5:1999. |
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吸出培养基，吸得越干净越好，直接加入1ml胰酶（T－25瓶），放到37度培养箱消化。是否需要用hanks 或 pbs之类得缓冲洗细胞并不绝对必要，我都没有洗，感觉应该洗一下更好，但也更麻烦并增加了污染得可能性。如果细胞长得太老，可以先加入1ml胰酶在细胞表面浸润一下就吸出来，然后再加入1ml胰酶消化。胰酶在使用前最好预热到37度。由于胰酶平时保存在4度，反复预热对胰酶的活性不好，我的经验是将胰酶进行分装，尽可能避免胰酶反复冷却加热。消化时间的选择要根据实际情况决定，BHK－21还是比较好消化
1、吸尽旧的培养液（因为我是养在培养皿)2、用D-HANKs清洗一次3、加入0.125%/0.02%EDTA的胰酶(盖满皿底即可)。肉眼观察即可，看见皿底有一层薄雾。4、加入完全培养基，以终止胰酶作用。5、轻轻反复吹打细胞。6、吸出一部分加入新的培养皿中。注意：1、可以用MEM或DMEM2、EDTA可使细胞分散3、BHK-21生长迅速
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BHK-21 [C-13]
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