KC-0629

NCI-H929-Cell-Line-(Not-for-sale)

20731

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Background of NCI-H929-Cell-Line-(Not-for-sale)

NCI-H929 cells are B lymphocytes isolated from a malignant effusion in a 62-year-old, White, female plasmacytoma myeloma patient.

Specifications

Catalog Number	KC-0629
Cell Line Name	NCI-H929-Cell-Line-(Not-for-sale)
Host Cell Line	NA
Description	Human multiple myeloma cell line, established from the pleural effusion of a 62-year-old white woman with myeloma (IgAkappa) at relapse
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	3D cell culture; Cancer research
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+0.05mM 2-mercaptoethanol
Selection Marker	NA
Morphology	Single, round to oval cells or clustered in small clumps in suspension
Subculture	Split saturated culture 1:2-1:3 every 3-4 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 50 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

Characterization

Cell Resuscitation

1. Prewarm culture medium (RPMI1640+10%FBS+0.05mM 2-mercaptoethanol)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:2-1:3 every 3-4 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1. Establishment and characterization of a human plasma cell myeloma culture having a rearranged cellular myc proto-oncogene. Blood 67:1542-1549(1986)