-Detailed Information [JCRB0085]-

JCRB0085 HL60

Cell information

Important Notice(s)

HL60細胞の解凍と培養について

ヒト血球系細胞株は一般的に解凍からの増殖の立ち上がりが悪く、HL60細胞ではこれが特に顕著です。場合によっては解凍時のショックで死滅してしまう場合があります。念のため以下の点に留意してみて下さい。

凍結状態の細胞は-80度以上の温度にさらされますと、それが短時間であっても急激に生存率が低下します。このため、解凍速度がかなり生存率に影響します。解凍は37度程度の微温中で振蕩することに急速に行って下さい。解凍までに1-2分が目安です。解凍後のDMSOを除くための遠心は1000 rpm　一回にとどめてください。

HL60細胞株の解凍からの増殖がうまくゆくかどうかは、細胞密度に依存することが多いです。これは、細胞自身が放出するautocrine factorの影響と考えられます。そこで、解凍からの最初の培養においては、局所的に細胞密度を高くしてやる、たとえば5mlの培地で培養を開始し、フラスコを傾けて角に細胞が集まるようにすることにより、増殖を促す効果が期待されます。バンクでは平坦な状態で解凍後の生存・増殖を確認はしていますが、念のためお試しください。

また、血清濃度を一時的に20%にしてやることも効果があります。同様の理由で、頻繁な培地交換はむしろautocrine factorの希釈につながり、立ち上がりが悪くなる原因となり得ます。解凍時にDMSOを除くための遠心を一度行ったら、培養開始後は増殖が確認されるまで培地交換はむしろ行わない方がよい結果が得られます。死細胞が増えたとしても増殖を始めるまでは培地交換も頻繁には行わず、培地の色が黄色になるまで放置して下さい。また、細胞が対数増殖期に入るまで、１週間以上かかることもよくあります。凍結直後の継代は、細胞がある程度フラスコを覆うまで待ち、様子を見ながら1:1くらいから始めて下さい。継代時には、それまでに培養されていた培地をそのまま新鮮な培地に対して希釈してください。

解凍時・再培養時には高pHが強いダメージを与えます。基礎培地のRPMI1640は、pHが高くなりやすい傾向がありますのでインキュベーター中の培地のpHに注意してください。もしインキュベーター中でのpHが高めになるようでしたら、予めpHを低めにRPMI1640を調製してください。

抗生物質としては、100 units/ml penicillin & 100 μg/ml streptomycin あるいは50 μg/ml kanamycin の使用は問題ないと思います。しかし、もしFungison (amphotericin B)やgentamaicinを使われている場合は特に前者の使用を避けてください。影響は少ないと思いますが、いずれの細胞に対しても増殖阻害的に働きます。



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The cultivation of HL60: Tips for recovery.

[The cultivation of HL60: Tips for recovery after thawing.]

Human haematopoietic cell lines often fall into growth problem after thawing. This tendancy is prominent for HL60 cell line, and sometimes the most of cells will die due to the shock by thawing. To improve the condition of cells, please take care the following matters.

The viability of frozen cells rapidly decrease by the exposure to the temperature higher than -65C even the exposure time is short. Therefore the frozen vial should be immediately thaw after taking from dry ice, or freezer/liquid nitorgen tank. The thawing should be made rapidly by immersing the vial to the 37 C water bath within 1-2 min. It is essential to remove DMSO after thawing by centrifugation (1000 rpm, 3-5 min). Repeated centrifugation is not necessary.

The growth of HL60 cell line tend to depend on high cell density. This is because the cell line itself secretes the growth factors and survival factors in the culture media. The most simple method will be slant culture. The flask is slanted so as to achieve the local high cell density in the bottom of flask corner.

The autocrine factor(s) may be substituted by serum. Therefore, please increase serum concentration in culture medium to 20% until the cells stably grow.

By the same reason, excessive dilution or too frequent medium renewal may become rather harmful to cells. Simply add the medium to the old medium with cells.

The pH of RPMI1640 medium tends to become alkaline. The alkaline pH is very harmful for HL60. The media color indicated by phenol red should be orange. If the medium color is pink or purple, it is not suitable. If the pH of culture medium in CO2 incubator is too high, please prepare the RPMI1640 medium to somewhat lower pH.

The addition of 100 units/ml penicillin & 100 ug/ml streptomycin, or 50 ug/ml kanamycin is no problem. However, if you use amphotericin B + gentamycin, please avoid to use (especially amphotericin B). These sometimes inhibit the growth of cells.

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Cell type：general cells (View Pricing Information)

JCRB No.	JCRB0085	Cell Name	HL60
Profile	Promyelocytic cell line differentiated to neutrophils or macropahges by tumor promoters, vitamne D3 or cytekines. The cell line can be used for the research of the differenciation induction or analysis of some oncogenes.	Other Name	HL-60
Animal	human	Strain
Genus	Homo	Species	sapiens
Sex	F	Age	36
Identity	available	Tissue for Primary Cancer	hemo-lymphocytic
Case history	acute promyelotic leukemia	Metastasis
Tissue Metastasized		Genetics	APL
Life Span	infinite	Crisis PDL
Morphology	lymphocyte-like	Character	DMSO,butyl.,HX.,TPA,act.D,& R.A. ind. differentiat
Classify	tumor	Established by	Collins,S.J. et al.
Registered by	Hozumi,M.	Regulation for Distribution	free
Comment	Doubling time is 40hr. Growth is not so good from thawing to 1 month. Inoculate about 10^6 cells to 60mm dish when thawing. STR-PCR indicated the HL60 is same as HL60(S) (JCRB0163),	Year	1986
Medium	RPMI1640 medium with 20% fetal calf serum.	Methods for Passages	Simple dilution.
Cell Number on Passage		Race	Cocasian
CO2 Conc.	5 %	Tissue Sampling	peripheral blood
Tissue Type

Detection of virus genome fragment by Real-time PCR
Detected DNA Virus	tested				Detected RNA Virus	tested
	CMV	-	parvoB19	-		HCV	-	HTLV-1	-
	EBV	-	HBV	-		HIV-1	-	HTLV-2	-
	HHV6	-	HTLV-1	-		HIV-2	-	HAV	-
	HHV7	-	HTLV-2	-		-/negative. +/positive. nt/not tested. (positive (+) does not immediately mean the production of infectious viral particles.)
	BKV	-	HIV-1	-
	JCV	-	HIV-2	-
	ADV	-	HPV18	-
Notes

Link
Cellosaurus(ExPASy) Cellosaurus (ExPASy) is developed by Amos Bairoch of the CALIPHO group at the SIB - Swiss Institute of Bioinformatics as part of the neXtProt project. Cellosaurus is a knowledge database of cell lines with various information summarized.	CVCL_0002

Reference
Pubmed id:10470292	Contribution of the nucleoside transport system to doxorubicin transport in HL60 cells but not in mononuclear cells. Nagasawa K,Fumihara T,Ohnishi N,Yokoyama T Jpn J Cancer Res. 1999 Jul;90(7):781-7
Pubmed id:6192693	Fundamentals of chemotherapy of myeloid leukemia by induction of leukemia cell differentiation. Hozumi M Adv Cancer Res. 1983;38():121-69
Pubmed id:288488	Characterization of the continuous, differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia. Gallagher R,Collins S,Trujillo J,McCredie K,Ahearn M,Tsai S,Metzgar R,Aulakh G,Ting R,Ruscetti F,Gallo R Blood. 1979 Sep;54(3):713-33
Pubmed id:271272	Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Collins SJ,Gallo RC,Gallagher RE Nature. 1977 Nov 24;270(5635):347-9

Images

LOT Information

08192010
04282009
11182005
11062001
042197
051496
031495
073093
080493
080593
091792
042092
062091
090790
051590
101189
080687
043097
02082016
02152018
04092019
07072022

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	08192010	Lot Specification	distribution
Medium	RPMI1640 medium with 10% fetal bovine serum (GIBCO 10099). Use 20% FBS until cell growth becomes log phase.	Temperature	37 C
Cell Density at Seeding	1 - 3 x 10^5 cells/ml	Methods for Passages	dilution (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです)
Doubling Time		Cell Number in Vial (cells/1ml)	1.1 x 10^6
Viability at cell freezing (%)	79	Antibiotics Used	free
Passage Number	Unknown (10 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	Confirmed as human by NP, G6PD (type B), MD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI1640	CO2 Conc.	5%
Viability immediately after thawing (%)	79	Additional information

Cell No.	JCRB0085	Cell Name
LOT No.	04282009	Lot Specification	distribution
Medium	RPMI1640 medium with 10% fetal bovine serum (GIBCO 10099). Use 20% FBS until cell growth becomes log phase.	Temperature	37 C
Cell Density at Seeding	1 - 3 x 10^5 cells/ml	Methods for Passages	dilution (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです)
Doubling Time		Cell Number in Vial (cells/1ml)	4.8 x 10^6
Viability at cell freezing (%)	92.5	Antibiotics Used	free
Passage Number	Unknown (12 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	Confirmed as human by NP, G6PD (type B), MD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI 1640	CO2 Conc.	5%
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name
LOT No.	11182005	Lot Specification	distribution
Medium	RPMI1640 medium with 10% fetal bovine serum (FBS lot; GIBCO 4218333S). Use 20% FBS until cells growth becomes log phase.	Temperature	37 C
Cell Density at Seeding	1.5 - 3 x 10^5 cells/ml.	Methods for Passages	dilution (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです)
Doubling Time	30-40 hrs	Cell Number in Vial (cells/1ml)	9.3 x 10^5
Viability at cell freezing (%)	92	Antibiotics Used	free
Passage Number	Unknown (12 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI1640	CO2 Conc.	5%
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name
LOT No.	11062001	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum(非働化を推奨。対数増殖期は10%で培養可能)	Temperature	37 C
Cell Density at Seeding	1.5-3.0 x 10^5 cells/ml	Methods for Passages	dilution. 解凍直後は5mlの培地, FBS 20%, フラスコを傾けて培養すると立上りが良好
Doubling Time	D.T. = ca 40 hrs	Cell Number in Vial (cells/1ml)	3.0 x 10^6
Viability at cell freezing (%)	87.0	Antibiotics Used	free
Passage Number	P12*	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS-RPMI1640	CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	042197	Lot Specification	distribution
Medium	RPMI1640 medium with 10% or 20% heat inactivated fetal bovine serum (Cell Culture T.1604309).	Temperature	37 C
Cell Density at Seeding	2x10^5 cells/ml	Methods for Passages	Simple dilution, because of a suspension culture.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	1.93x10^6
Viability at cell freezing (%)	96.3	Antibiotics Used	free
Passage Number	P10*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	Under investigation.
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing	Culture medium with 5% DMSO.	CO2 Conc.
Viability immediately after thawing (%)		Additional information

Images

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	051496	Lot Specification	distribution
Medium		Temperature
Cell Density at Seeding		Methods for Passages
Doubling Time		Cell Number in Vial (cells/1ml)
Viability at cell freezing (%)		Antibiotics Used
Passage Number		PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Images

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	031495	Lot Specification	distribution
Medium	RPMI1640 medium with 20% heat inactivated fetal bovine serum (JRS.DO10400)	Temperature	37 C
Cell Density at Seeding	2-5x10^5 cells/ml	Methods for Passages	dilution
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.9x10^6
Viability at cell freezing (%)	92.8	Antibiotics Used	free
Passage Number	P7*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	073093	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum (Hyclone lot.1151031)	Temperature	37 C
Cell Density at Seeding	2-5x10^5 cells/ml	Methods for Passages	Cells are simply diluted with fresh medium.
Doubling Time	86.6hr->51.4hr	Cell Number in Vial (cells/1ml)	3.6x10^6
Viability at cell freezing (%)	81.3	Antibiotics Used	free
Passage Number	P13*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	LDH,G6PD,NP G6PD=type B
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	080493	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum (JRH Biosciences Lot.1B1062)	Temperature	37 C
Cell Density at Seeding	2-3x10^5 cells/ml	Methods for Passages	Cells are simply diluted by the fresh medium.
Doubling Time	Td=45.2-52.2hrs	Cell Number in Vial (cells/1ml)	4.3x10^6
Viability at cell freezing (%)	78.3	Antibiotics Used	free
Passage Number	P14*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	080593	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum (JRH Biosciences, lot.1B1062)	Temperature	37 C
Cell Density at Seeding	2-3x10^5 cells/ml	Methods for Passages	Cells are diluted with fresh medium.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.8x10^6
Viability at cell freezing (%)	84.5	Antibiotics Used	free
Passage Number	P14*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	091792	Lot Specification	distribution
Medium	RPMI1640 medium (081792) with 10% fetal bovine serum (Hyclone 11151031)	Temperature	37 C
Cell Density at Seeding	1.1-1.5x10^5 cells/ml	Methods for Passages	Simple dilution
Doubling Time	NT	Cell Number in Vial (cells/1ml)	1.37x10^6
Viability at cell freezing (%)	94.9	Antibiotics Used	free
Passage Number	P7* =correct.	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	M=46	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing	Culture medium with 5% DMSO.	CO2 Conc.
Viability immediately after thawing (%)		Additional information

Images

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	042092	Lot Specification	distribution
Medium	RPMI1640(lot.021992,041592)+10% FBS(lot.Cell Cult.Lab. 0099-01)	Temperature	37 C
Cell Density at Seeding	3x10^4 cells/sq.cm	Methods for Passages	dilution
Doubling Time	NT	Cell Number in Vial (cells/1ml)	8.9x10^6
Viability at cell freezing (%)	86.9	Antibiotics Used	free
Passage Number	P7*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	062091	Lot Specification	distribution
Medium	RPMI1640 + 10 % FBS	Temperature	37
Cell Density at Seeding	1-2x10^5 cells/ml	Methods for Passages	Dilution subculture
Doubling Time	NT	Cell Number in Vial (cells/1ml)	1x10^6
Viability at cell freezing (%)	97.4	Antibiotics Used	free
Passage Number	P6*	PDL	NT
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode	NT	Chromosome Information	NT
Surface Antigen	NT	DNA Profile (STR)
Adhesion	No	Exoteric Gene	NT
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	090790	Lot Specification	distribution
Medium	RPMI1640 + FCS 10%	Temperature	37
Cell Density at Seeding	1-2x10^5 cells/ml	Methods for Passages	Dilute by fresh medium, and subculture every 6-7 days.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	5.0x10^6
Viability at cell freezing (%)	NT	Antibiotics Used	free
Passage Number	P7	PDL
Sterility: MYCOPLASMA	NT	Sterility: BACTERIA	NT
Sterility: FUNGI	NT	Isozyme Analysis	NT
Chromosome Mode		Chromosome Information	NT
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	051590	Lot Specification	distribution
Medium	RPMI1640 + FCS 10%	Temperature	37
Cell Density at Seeding	1-2x10^5 cells/ml	Methods for Passages	Dilute by fresh medium, and subculture every 6-7 days.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.5x10^6
Viability at cell freezing (%)	NT	Antibiotics Used	free
Passage Number	P8	PDL
Sterility: MYCOPLASMA	NT	Sterility: BACTERIA	NT
Sterility: FUNGI	NT	Isozyme Analysis	NT
Chromosome Mode		Chromosome Information	NT
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	101189	Lot Specification	distribution
Medium	RPMI1640 + FCS 10%	Temperature	37
Cell Density at Seeding	1-2x10^5 cells/ml	Methods for Passages	Dilute by fresh medium, and subculture every 6-7 days.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	2.7x10^6
Viability at cell freezing (%)	NT	Antibiotics Used	free
Passage Number	P7*	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode		Chromosome Information	NT
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	080687	Lot Specification	distribution
Medium	RPMI1640 + FCS 10%	Temperature	37
Cell Density at Seeding	1-2x10^5 cells/ml	Methods for Passages	Dilute by fresh medium, and subculture every 6-7 days.
Doubling Time	NT	Cell Number in Vial (cells/1ml)	9x10^5
Viability at cell freezing (%)	NT	Antibiotics Used	free
Passage Number	P6*	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	G6PD,LD,NP
Chromosome Mode		Chromosome Information	NT
Surface Antigen		DNA Profile (STR)
Adhesion		Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name
LOT No.	043097	Lot Specification	distribution
Medium	RPMI1640 with 20% fetal bovine serum(FBSは熱非働化を推奨)。対数増殖期にはFBS10%にて培養可能。	Temperature	37 C
Cell Density at Seeding	ca 2 x 10^5 cells/ml	Methods for Passages	希釈。解凍時は5mlの培地から御培養下さい。フラスコを傾け細胞を寄集めた状態で培養すると立上りが良好。
Doubling Time		Cell Number in Vial (cells/1ml)	1.9 x 10^6
Viability at cell freezing (%)	93.0	Antibiotics Used	free
Passage Number	P13*	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	confirmed as human by NP, G6PD (type B), MD.
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing		CO2 Conc.
Viability immediately after thawing (%)		Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	02082016	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum (FBS; GIBCO Cat. # 10091).	Temperature	37 C
Cell Density at Seeding	2.6 - 2.8 x 10^5 cells/mL	Methods for Passages	Simple dilution. (Slant culture to achieve local high cell density on first seeding)
Doubling Time	approx. 31 hrs.	Cell Number in Vial (cells/1ml)	3.6 x 10^6
Viability at cell freezing (%)	84	Antibiotics Used	free
Passage Number	Unknown (11 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis	NT
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI1640	CO2 Conc.	5%
Viability immediately after thawing (%)	84	Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	02152018	Lot Specification	distribution
Medium	RPMI1640 medium with 20% fetal bovine serum (FBS; Sigma Cat. # 172012).	Temperature	37 C
Cell Density at Seeding	0.7 - 1.3 x 10^5 cells/mL	Methods for Passages	Simple dilution (suspension culture). (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです　To improve first growth, the flask should be slant slightly to gather cells to the corner. )
Doubling Time	approx. 27 hrs.	Cell Number in Vial (cells/1ml)	4.9 x 10^6
Viability at cell freezing (%)	90.1	Antibiotics Used	free
Passage Number	Unknown (15 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI1640	CO2 Conc.	5%
Viability immediately after thawing (%)	90.1	Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	04092019	Lot Specification	distribution
Medium	RPMI 1640 medium with 20% fetal bovine serum (FBS; Sigma Cat. # 172012).	Temperature	37 C
Cell Density at Seeding	approx. 1.5 x 10^5 cells/mL	Methods for Passages	Simple dilution (suspension culture). (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです　To improve first growth, the flask should be slant slightly to gather cells to the corner. )
Doubling Time	approx. 32 hrs.	Cell Number in Vial (cells/1ml)	2.8 x 10^6
Viability at cell freezing (%)	91.7	Antibiotics Used	free
Passage Number	Unknown (10 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI 1640	CO2 Conc.	5%
Viability immediately after thawing (%)	83.0	Additional information

Cell No.	JCRB0085	Cell Name	HL60
LOT No.	07072022	Lot Specification	distribution
Medium	RPMI 1640 medium with 20% fetal bovine serum (FBS; Biowest Cat. # S1820-500).	Temperature	37 C
Cell Density at Seeding	1 - 3 x 10^5 cells/mL	Methods for Passages	Simple dilution (suspension culture). (解凍時はフラスコを傾けて細胞を角に集めて培養すると増殖の立ち上がりが早いです　To improve first growth, the flask should be slant slightly to gather cells to the corner. )
Doubling Time	approx. 34 hrs.	Cell Number in Vial (cells/1ml)	4.9 x 10^6
Viability at cell freezing (%)	96	Antibiotics Used	free
Passage Number	Unknown (12 at bank)	PDL
Sterility: MYCOPLASMA	-	Sterility: BACTERIA	-
Sterility: FUNGI	-	Isozyme Analysis
Chromosome Mode		Chromosome Information
Surface Antigen		DNA Profile (STR)	D5S818:12 D13S317:8,11 D7S820:11,12 D16S539:11 VWA:16 TH01:7,8 AM:X TPOX:8,11 CSF1PO:13,14
Adhesion	No	Exoteric Gene
Medium for Freezing	10% DMSO, 20% FBS - RPMI1640	CO2 Conc.	5%
Viability immediately after thawing (%)	91	Additional information

JCRB Cell Bank

JCRB0085 HL60

Cell information

LOT Information