HBL-100 Cells
General information
Description | The epithelial cell line HBL-100 has been derived by E.V. Gaffney and associates from the milk of a nursing mother and obtained 3 days after delivery. Although there was no evidence of a breast lesion in the milk donor, and the patient had no family history of breast cancer, the karyotype of the recovered cells was abnormal as early as passage 7. This line was able to synthesize a small amount of lactose and would respond to prolactin or estrogen by producing increased amounts of casein. Electron micrographs revealed microvilli, tonofibrils and desmosomes. Problematic cell line: Misidentified. Presence of a Y chromosome in cell line that was thought to be of female origin (Yoshino et al. 2006. Capes-Davies, 2010). Originally thought to originate from a casein-producing breast cell line. In addition contains SV40 genomic sequence while the cell line was deemed to be spontaneously immortalized. |
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Organism | Human |
Tissue | Breast |
Disease | Carcinoma |
Synonyms | HBL 100, HBL100 |
Characteristics
Age | 27 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | HBL-100 (Cytion catalog number 300178) |
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Biosafety level | 1 |
Expression / Mutation
Antigen expression | HLA A1, A10, A11, B7, B8 |
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Isoenzymes | G6PD, B, PGM1, 1, PGM3, 2, ES-D, 1, Me-2, 0, GLO-1, 2, AK-1, 1-2, Phenotype Frequency Product: 0.0008 |
Tumorigenic | Yes, in nude mice. At passage levels below 35 the line is not tumorigenic in nude mice, but forms colonies in soft agar. Tumorigenicity has been reported to increase above passage 35. |
Viruses | The cells contain a tamdemly integrated SV40 genome it has been reported that they may contain a type D retrovirus that is similar or identical to Mason-Pfizer monkey virus (MPMV). |
Reverse transcriptase | Positive |
Ploidy status | Aneuploid |
Karyotype | The stemline chromosome number is near triploid with the modal number of 67 chromosomes, and the 2S component occurring at 0.6%. Most chromosome complements consist of about 39 normal and 28 marker chromosomes. Markers such as 2q, 11q+, 11q, t(2q.12), t(2q.5q?), t(6p?.16), 16pt and many others are common to most metaphases. Normal chromosomes 11, 14, 15 and 16 are absent. 2, 12, 17 and 19 are monosomic, and the x is disomic. DNA profiling for amelogenin, a sex-chromosome-specific PCR assay that can distinguish x chromosome-specific products from Y chromosome-specific products revealed the presence of Y chromosomes in this cell line of putative female origin. Confirmation of the general findings was accomplished by QM staining, C-banding, and FISH, with a whole chromosome paint probe to the human Y chromosome. |
Handling
Culture Medium | McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:2 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,y
CSF1PO: 10
D13S317: 12
D16S539: 9,12
D5S818: 11,12
D7S820: 8,12
TH01: 6,8
TPOX: 8
vWA: 16
D3S1358: 14,16
D21S11: 28,30
D18S51: 16
Penta E: 7
Penta D: 12
D8S1179: 12,15
FGA: 25
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HLA alleles |
A*: 01:01:01, 02:01:01
B*: 08:01:01, 40:01:02
C*: 03:04:01, 07:01:01
DRB1*: 03:01:01, 15:01:01
DQA1*: 01:02:01, 05:01:01
DQB1*: 02:01:01, 06:02:01
DPB1*: 04:01:01
E: 01:01, 01:03
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