Cells are propagated from the stock, deposit
Passage Number, P5*
Cell culture was started without cloning from the previous stock.
Date cell culture started:07/16/2004
Concentration of cells in an ampule: 3.2 x 10^6 cell/ml
Viability under microscope (%): 62.9
Antibiotics used :Free. MC210 temporary added to remove mycoplasmas.
No contamination with bacteria and fungi but cells were found to be contaminated with mycoplasmas. Decontamination under process.
Anchorage dependency: Yes

Culture medium: DM201 medium. Serum free.
Freezing medium: 1 to 1 mixture of the culture medium and the FM-1 medium containing 10% DMSO.
Passage method: Cells were harvested by pipett or scraper. Subculture every 8-14 days and medium change every 2-4 days.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 4.5 x 10^4 cells/sq.cm.
Additional comments: Mycoplasmas tested by PCR(Ozawa,Y.) method. Once this lot was eliminated mycoplasmas by MC210. Later appeared again.

[ Additional Data ]

Cells(1), Cells(2), Cells(3), Mycoplasma test, Mycoplasma (PCR), Freezing process, Others(1),

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(Date downloaded: 2005-01-28)

Cells are propagated from the stock, deposit
Passage Number, P5*
Cell culture was started without cloning from the previous stock.
Date cell culture started:07/16/2004
Concentration of cells in an ampule: 2.3 x 10^6 cell/ml
Viability under microscope (%): 62.5
Antibiotics used :Free. MC210 temporary added to remove mycoplasmas.
No contamination with bacteria and fungi but cells were found to be contaminated with mycoplasmas. Decontamination under process.
Tested Isoenzymes, G6PD(typeB), NP, LDH examined. Human not HeLa..
Anchorage dependency: Yes

Culture medium: DM201 medium. Serum free.
Freezing medium: 1 to 1 mixture of the culture medium and the FM-1 medium containing 10% DMSO.
Passage method: Cells were harvested by pipett or scraper. Subculture every 8-14 days and medium change every 2-4 days.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 4.5 x 10^4 cells/sq.cm.
Additional comments: Mycoplasmas tested by PCR(Ozawa,Y.) method. While once eliminated mycoplasmas by MC210, later they were appeared again.

[ Additional Data ]

Cells(1), Cells(2), Cells(3), Mycoplasma test, Mycoplasma (PCR), Isozyme analysis, Freezing process, Others(1),

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(Date downloaded: 2005-01-28)

Cells are propagated from the stock, 08052004
Passage Number, P13*
Cell culture was started without cloning from the previous stock.
Date cell culture started:11/15/2004
Concentration of cells in an ampule: 1.1 x 10^6 cell/ml
Viability under microscope (%): 86.2
Antibiotics used :BMC-1&2 temporary.
Anchorage dependency: Yes
Cell Identification: available

Culture medium: DM201
Freezing medium: One to one mix of the culture medium and FM-1 medium containing 10% DMSO finally.
Passage method: Cells are harvested by pipetting with EDTA-PBS. Subculture every 4-7 days and medium change every 2-4 days.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 3.4 x 10^4 cells/sq.cm.
Additional comments: Mycoplasmas tested by staining method.

[ Additional Data ]

Cells(1), Cells(2), Cells(3), Mycoplasma test, STR-PCR(1), STR-PCR(2), Freezing process,

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(Date downloaded: 2005-01-28)

Cells are propagated from the stock, 08052004
Passage Number, P13*
Cell culture was started without cloning from the previous stock.
Date cell culture started:11/15/2004
Concentration of cells in an ampule: 1.7 x 10^6 cell/ml
Viability under microscope (%): 88.2
Antibiotics used :BMC-1 temporary.
Anchorage dependency: Yes
Cell Identification: available

Culture medium: DM201, serum free medium.
Freezing medium: One to one mix of DM201 and FM-1 medium containing 10% DMSO finally.
Passage method: Cells were harvested by pipetting with EDTA-PBS. Subculture every 4-7 days. Medium change every 2-4 days.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 3.4 x 10^4/sq.cm.
Additional comments: Mycoplasmas tested by staining method. It is hard to conclude only by this experiment.

[ Additional Data ]

Cells(1), Cells(2), Cells(3), STR-PCR(1), STR-PCR(2), Freezing process,

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(Date downloaded: 2005-01-28)

Cells are propagated from the stock, 08052004
Passage Number, P8*
Cell culture was started without cloning from the previous stock.
Date cell culture started:11/15/2004
Concentration of cells in an ampule: 2.8 x 10^6 cell/ml
Viability under microscope (%): 94.0
Antibiotics used :MC201 temporary
No contamination with bacteria and fungi but cells were found to be contaminated with mycoplasmas. Decontamination under process.
Anchorage dependency: Yes
Cell Identification: available

Culture medium: DM201 medium, serum free
Freezing medium: 1/1 mixture of DM210 and FM-1 containing 10% DMSO finally.
Passage method: Cells harvested by pipetting with EDTA-PBS.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 3.4 x 10^4 cells/sq.cm.
Comments: This lot should be discarded.

[ Additional Data ]

STR-PCR(1), STR-PCR(2),

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(Date downloaded: 2005-01-28)

Cells are propagated from the stock, 08052004
Passage Number, P9*
Cell culture was started without cloning from the previous stock.
Date cell culture started:11/15/2004
Concentration of cells in an ampule: 1.8 x 10^6 cell/ml
Viability under microscope (%): 88.6
Antibiotics used :free
Anchorage dependency: Yes

Culture medium: DM201 serum free medium.
Freezing medium: One to one mix of the DM201 and FM-1 containing 10% DMSO finally.
Passage method: Cells were harvested by pipetting with EDTA-PBS. Subculture once a week and medium change every 2-4 days.
Growth temperature: 37 C
CO₂ concentration: 5 %
Cell No. at passage: 3.4 x 10^4 cells/sq.cm.
Additional comments: Mycoplasmas tested by staining method.

[ Additional Data ]

Cells(1), Cells(2), Cells(3), Mycoplasma test, Freezing process,

---

(Date downloaded: 2005-01-28)