This kit contains Bone marrow stromal cells including undifferentiated mesenchymal stem cells.

Osteogenesis Culture Kit


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Background

There are hematopoietic stem cells and bone marrow stromal cells in bone marrow. Bone marrow stromal cells contain undifferentiated mesenchymal stem cells that can differentiate into a variety of cell types such as osteoblasts chondrocytes adipocytes and so on. Osteogenesis Culture Kit (Mouse) (PMC-OGC11-COS) contains cryopreserved cells isolated from mouse bone marrow and two types of culture medium.
The cells in this product can be grown using Growth Medium (Code No. PMC- OGCMG-COS), and then can be differentiated into mature osteoblasts, which form calcified nodules, using Culture Medium (Mouse) (Code No. PMC- OGCMO-COS).

Fig. 1 Morphology of cultured cells
A: Day 0
B: Confluent
C: 3 weeks after medium is changed into “Osteogenesis Culture Medium for Osteogenesis Culture kit” (35 mm dish)

Kit component

Product Name Size Quantity Storage Conditions Stability
Bone marrow stromal cell, ICR Mouse 1x 106 cells 1 Liquid Nitrogen 1 years
Growth medium*
(PMC-OGCMG-COS)
125 mL 1 -20°C Freezer 6 months
4°C 3 months
Culture medium**
(PMC-OGCMO-COS)
250 mL 1 -20°C Freezer 6 months
4°C 3 months

Components of Media Components of Media:
Growth medium for Osteogenesis Culture kit is sterile, liquid basal medium (α-MEM) which contain contains 10 % FBS, 10 units/ml penicillin and 10 μg/ml streptomycin.
** Culture medium for Osteogenesis Culture kit (Mouse) (Code No. PMC- OGCMO-COS) is sterile, liquid basal medium (α-MEM) which contain contains 10 % FBS, 10 units/ml penicillin, 10 μg/ml streptomycin, dexamethasone, ascorbic acid and β-Glycerophosphate.

Protocol : Cultured with the 24-wellplate

1. Thaw the Growth medium in a 37 C water bath with gentle shaking.

2. Quickly place Osteogenesis Cell vial in a 37°C water bath until the contents are thawed.
   * Immediately transfer cryovials from dry ice to liquid nitrogen upon receiving and keep the cells in liquid nitrogen until cell culture needed for experiments. It is recommended to use osteoblast cell vial immediately upon receiving.

3. Transfer thawed cells into a 15 mL centrifuge tube containing 10 mL of Growth medium and mix gently.

4. Centrifuge at 4 C at 600 x g for 5 minutes.

5. After removing the supernatant, re-suspend cells in 10 mL of Growth medium and centrifuge for 5 minutes at 4 C at 600 x g for 5 minutes.

6. After removing the supernatant, re-suspend the cell pellet in approximately 5 mL of Growth medium.

7. Dispense 0.5 mL of cell suspension to each well of 24-well plate.
   * In the case of using other types of culture plate, adjust cell density to 2.5×104 cells/ cm2.

8. Change the medium in the following day, and every other day thereafter until the cells become confluent.
   * Cells become confluent within 3-4 days .

9. Once cells become confluent, replace the medium with "Osteogenesis Culture Medium to promote the formation of calcified nodules.

10. Change the medium every 2 or 3 days.
   * Approximately 4-7 days after medium is changed into "Osteogenesis Culture for Osteogenesis Culture kit", cells whiten. Adipocytes appear from a part of cells.

Application Example

Cells are cultured according to protocol and subjected to activity staining of alkaline phosphatase or calcified nodules. The activity of alkaline phosphatase is detected by Alkaline Phosphatase Staining kit (Code: PMC-AK20-COS) and the calcium deposit is detected by Calcified nodule Staining kit (Code: PMC-AK21-COS).

References

> SL Cheng et al., Differentiation of human bone marrow osteogenic stromal cells in vitro: induction of the osteoblast phenotype by dexamethasone. Endocrinology, 134(1) p277-286 (1994)


> Ohgushi et al., In vitro bone formation by rat marrow cell culture. Journal of Biomedical Materials Research Part A, 32(3) p333-340 (1996)

Product List

Product Name Cat# Quantity Price

Osteogenesis Culture kit, Mouse DataSheet

PMC-OGC11-COS

1 VIAL ¥114,000
$1520

Osteogenesis Growth Medium, Mouse DataSheet

PMC-OGCMG-COS

250 ML ¥31,200
$416

Osteogenesis Culture Medium, Mouse DataSheet

PMC-OGCMO-COS

250 ML ¥31,200
$416

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.


“Osteogenesis Culture Kit” also belongs to the following categories.