KBM ADSC-1 (Adipose-Derived Stem Cell Culture Medium)

Cosmo Bio LTD

Catalog No: KJN-16030020

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Adipose-derived stem cell culture medium KBM ADSC-1&2

Long-term culture is possible while maintaining undifferentiated state
Manufacturer: Kohjin Bio Co., Ltd.

This product is a culture medium suitable for growing adipose-derived stem cells while maintaining their undifferentiation.
※The maintenance of undifferentiation is confirmed by the fact that cells cultured in this medium differentiate into adipocytes,　osteoblasts and chondrocytes, and by their surface markers.※

KBM ADSC-1&2, a culture medium for adipose-derived stem cells, enables long-term culture  while maintaining the undifferentiated state of adipose-derived stem cells (ADSC)  ^{(*) .}
KBM ADSC-1  is a ready-to-use medium containing 5% FBS. We  also sell  KBM ADSC-2,
which is KBM ADSC-1 without FBS .
^(*)  Differentiation into fat, bone, and cartilage confirmed

Intended use

It can be used for research on differentiation induction (fat, bone, cartilage, etc.) and lipid production.

Culture examples

Example of culture using KBM ADSC-1

Figure 1 Growth curve during subculture 
Figure 2. Changes in PDT (population doubling time) during subculture

Table 1. Percentage of various surface markers during subculture

(%)	P5	P6	P7
CD29	100	100	100
CD44	100	100	100
CD73	100	100	100
CD90	99.6	99.5	99.2
CD105	99.5	99.5	97.3

Culture conditions

• Cells: ADSC (commercially available P1 frozen cells)
• Container: 6-well cell culture plate (Corning, product number: 3516)
• Culture:
  4.75 × 104  ^cells  were seeded in a 6-well cell culture plate. On day 3 of culture, the cells were counted, subcultured, and analyzed for surface markers by flow cytometry.

   

Figure 3
(a) P3 ADSCs on day 3 of culture
(b) P5 cells on day 7 after induction of adipogenic differentiation (Oil Red O staining)
(c) P5 cells on day 21 after induction of osteogenic differentiation (Alizarin Red S staining)
(d) P3 cells on day 28 after induction of chondrogenic differentiation (Alcian Blue staining)

Culture conditions

• Cells: ADSCs (commercially available P1 frozen cells) cultured for 2 passages in KBM ADSC-1.
• Container: 24-well cell culture plate (FALCON, product number: 353047)
• culture:
  • 6.25 x 104  ^fat  cells  were seeded on a 24-well cell culture plate. On the first day of culture in culture medium (D-MEM(H) + 10% FBS + 1mM Na pyruvate),
    differentiation induction was started with KBM Adipogenic-DM (tentative), the medium was changed every 2-3 days, and on the 7th day of culture, staining was done with Oil Red O.
  • 6.25 x 104  ^bone  cells  were seeded in a 24-well cell culture plate. On the first day of culture in culture medium (D-MEM(H) + 10% FBS),
    differentiation induction was started with KBM Osteogenic-DM1 (tentative), and the medium was changed every 2 to 3 days. On the 21st day of culture, the cells were stained with Alizarin Red S.
  • 6.25 x 104  ^cells  of cartilage  were seeded onto a 24-well cell culture plate. On the first day of culture in culture medium (D-MEM(H) + 10% FBS),
    differentiation induction was started with a general cartilage differentiation induction medium, the medium was changed every 3 to 4 days, and on the 28th day of culture, staining was performed with Alcian Blue.

Culture example using KBM ADSC-2

Figure 4 Growth curve during subculture 
Figure 5. Changes in PDT (population doubling time) during subculture

Table 2. Percentage of various surface markers during subculture

(%)	Medium	P3	P4	P5	P7	P8
CD29	FBS	99.9	99.2	98.9	99.9	99.6
	HS	99.7	97.9	99.2	99.7	99.9
CD44	FBS	99.8	100	100	100	99.9
	HS	100	99.9	100	100	99.9
CD73	FBS	99.9	99.8	99.9	99.9	100
	HS	99.8	99.7	99.5	99.9	99.7
CD90	FBS	100	99.9	99.6	99	99.9
	HS	99.8	99.6	99.7	99.7	99.9
CD105	FBS	99.8	99.6	99.8	99.8	99.3
	HS	99.9	99.7	99.1	99.4	99.4

Culture conditions

• Cells: ADSC (commercially available P2 frozen cells)
• Container: 6-well cell culture plate (Corning, product number: 3516)
• Culture: 4.75 × 104  ^cells  were seeded in a 6-well cell culture plate. On the third day of culture, the cells were counted, subcultured, and analyzed for surface markers by flow cytometry.

   

Figure 6
(a) ADSC-2 + ADSCs on day 3 of P3 culture cultured with 5% FBS
(b) ADSC-2 + ADSCs on day 3 of P3 culture cultured with 5% HS
(c) HS P8 cells on day 7 after induction of adipogenic differentiation (Oil Red O staining)
(d) HS P8 cells on day 21 after induction of osteogenic differentiation (Alizarin Red S staining)

Culture conditions

• Cells: ADSCs (commercially available P2 frozen cells) cultured for 1-5 passages in KBM ADSC-2 + 5% FBS or KBM ADSC-2 + 5% HS
• Container: 24-well cell culture plate (FALCON, product number: 353047)
• culture:
  • 6.25 x 104  ^fat  cells  were seeded onto a 24-well cell culture plate. On the first day of culture in culture medium (D-MEM(H) + 10% FBS),
    differentiation induction was started with KBM Adipogenic-DM (tentative), and the medium was changed every 2-3 days. On the 7th day of culture, staining was performed with Oil Red O.
  • 6.25 x 104  ^bone  cells  were seeded in a 24-well cell culture plate.
    On the first day of culture in culture medium (D-MEM(H) + 10% FBS), differentiation induction was started with KBM Osteogenic-DM (tentative), and the medium was changed every 2 to 3 days. On the 21st day of culture, the cells were stained with Alizarin Red S.

Adipose-derived stem cell culture medium KBM ADSC-1 (5% FBM)

Product name	Catalog number	Size
KBM ADSC-1 (with FBS)	KJN-16030020	500ml

Adipose-derived stem cell culture medium KBM ADSC-2 (FBS-free)

Product name	Catalog number	Size
KBM ADSC-2 (w/o FBS)	KJN-16030030	500ml

Citations for KBM ADSC-1 (Adipose-Derived Stem Cell Culture Medium) – 4 Found

Nomura, Masateru; George, Joseph; Hashizume, Chieko; Saito, Takashi; Ueda, Yoshimichi; Ishigaki, Yasuhito; Tsuchishima, Mutsumi; Tsutsumi, Mikihiro. Surgical implantation of human adipose derived stem cells attenuates experimentally induced hepatic fibrosis in rats. Molecular Medicine (Cambridge, Mass.). 2022;28(1):143. 

Ishikura, Tomoko; Shiga, Hideaki; Nakamura, Yuka; Kanitani, Takako; Ishigaki, Yasuhito; Miwa, Takaki. Olfactory Regeneration with Nasally Administered Murine Adipose-Derived Stem Cells in Olfactory Epithelium Damaged Mice. Cells. 2023;12(5) 
- Comments: Loss of multipotency in adipose-derived stem cells after culture in temperature-responsive injectable polymer hydrogels. Polymer Journal 2022 https://www.nature.com/articles/s41428-022-00739-4

Kitajima, Hironori; Sakamoto, Takuya; Horie, Tetsuhiro; Kuwano, Ayane; Fuku, Atsushi; Taki, Yasuhiko; Nakamura, Yuka; Tanida, Ikuhiro; Sunami, Hiroshi; Hirata, Hiroaki; Tachi, Yoshiyuki; Yamamoto, Naoki; Iida, Yasuo; Ishigaki, Yasuhito; Yamada, Sohsuke; Shimodaira, Shigetaka; Shimizu, Yusuke; Ichiseki, Toru; Kaneuji, Ayumi; Osawa, Satoshi; Kawahara, Norio. Synovial Fluid Derived from Human Knee Osteoarthritis Increases the Viability of Human Adipose-Derived Stem Cells through Upregulation of FOSL1. Cells. 2023;12(2) 

Hasegawa, Kiko; Takenaka, Nobuyuki; Yamamoto, Maaya; Sakoda, Yoshiki; Aiba, Atsu; Satoh, Takaya. Regulation of De Novo Lipid Synthesis by the Small GTPase Rac1 in the Adipogenic Differentiation of Progenitor Cells from Mouse White Adipose Tissue. International Journal Of Molecular Sciences. 2023;24(5)