140-006-238.01
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Miltenyi Biotec B.V. & Co. KG
Friedrich-Ebert-Straße 68, 51429 Bergisch Gladbach, Germany
Phone +49 2204 8306-0, Fax +49 2204 85197
macsde@miltenyi.com
www.miltenyibiotec.com
Contents
1. Description
1.1 Background information
1.2 Applications
1.3 Reagent requirements
2. Protocol
2.1 Freezing of primary cells
2.2 Thawing of primary cells
2.3 Freezing of tissues
2.4 Thawing of tissues
1. Description
is product is for research use only.
Components 50 mL MACS Freezing Solution
Specications pH: 7.4−7.8
Contains 10% DMSO as cryoprotectant.
Storage Store protected from light at 2−8 °C. e expiration
date is indicated on the vial label.
1.1 Background information
e MACS Freezing Solution is a ready-to-use animal component-
free media formulation designed for the xeno- and serum-free
cryopreservation of primary cells and solid tissues, such as human
and mouse tumor samples, peripheral blood mononuclear cells
(PBMCs), and cells from dissociated solid tissues (e.g. cells from
dissociated tumors). Cells and tissues cryopreserved in MACS
Freezing Solution show high preservation of viability and good
recovery aer thawing.
1.2 Applications
● Cryopreservation of primary cells for downstream applications
(e.g. cell analysis, cell culture)
● Cyropreservation of solid tissues for downstream applications
(e.g. dissociation into single-cell suspensions)
1.3 Reagent requirements
● Cell culture medium
● Cryogenic vials
● Freezing container
● (Only for tissue pieces) MACS SmartStrainers (70 µm)
(#130-098-462)
● (Optional) MACS Tissue Dissociation Kits (e.g. Tumor
Dissociation Kit, human (# 130-095-929)) in combination with
the gentleMACS™ Octo Dissociator with Heaters (#130-096-
427) for the preparation of single-cell suspensions from solid
tissues
2. Protocol
▲ For the handling of the freezing container refer to the instructions
of the provider.
2.1 Freezing of primary cells
1. Determine cell number.
2. Centrifuge cell suspension at 300×g for 10 minutes. Aspirate
supernatant completely.
3. Resuspend cell pellet in MACS Freezing Solution to reach a
concentration of 10⁷ cells per mL.
4. Quickly transfer the cell suspension into cryogenic vials.
5. Place the vials in a freezing container and immediately store at
−80 °C.
6. Aer 24 hours transfer the cryogenic vials with the cells into a
liquid nitrogen tank for long-term storage.
2.2 Thawing of primary cells
▲ Work quickly to avoid loss of cells.
▲ Pre-heat water bath at 37 °C before taking cells out of liquid
nitrogen.
1. Take a vial with cells out of the liquid nitrogen tank.
2. Incubate the vial in a pre-heated water bath at 37 °C until only
a little lump of ice is le.
3. Quickly transfer cell suspension into a 15 mL conical tube and
dropwise add 5 mL of cell culture medium.
4. Wash cryogenic vial with 1 mL of cell culture medium and add
to conical tube.
5. Centrifuge cells for 5 minutes at 300×g. Aspirate supernatant
carefully.
6. Resuspend the cell pellet in appropriate buer according to
downstream application.
2.3 Freezing of tissues
1. Determine tissue weight.
2. Cut tissue into small pieces as recommended for downstream
application (e.g. tissue dissociation).
▲ Note: If using the MACS Tissue Dissociation Kits in combination with the
gentleMACS Octo Dissociator with Heaters for downstream tissue dissociation,
cut the tissue as indicated in the respective data sheet.
3. Transfer up to 200 mg of tissue pieces per 1 mL of capacity into
the cryogenic vials.
4. Add 1 mL of MACS Freezing Solution to up to 200 mg of tissue
pieces. For larger tissue amounts scale up accordingly.
5. Close the cryogenic vials and resuspend tissue pieces by
inverting the vial several times until pieces oat in MACS
Freezing Solution.
MACS® Freezing Solution
Order no. 130-129-552
Order no. 130-129-552
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Unless otherwise specically indicated, all Miltenyi Biotec products and services
are for research use only and not for diagnostic or therapeutic use.
140-006-238.01
6. Place the vials in a freezing container and immediately store at
−80 °C.
7. Aer 24 hours transfer the cryogenic vials with the tissue
pieces into a liquid nitrogen tank for long-term storage.
2.4 Thawing of tissues
▲ Work quickly to avoid loss of cells.
▲ Pre-heat water bath at 37 °C before taking tissues out of liquid
nitrogen.
1. Place a MACS SmartStrainer (70 µm) on a 50 mL conical tube.
2. Rinse the MACS SmartStrainer (70 µm) with 1 mL of cell
culture medium.
3. Take a vial with tissue pieces out of the liquid nitrogen tank.
4. Incubate the vial in a pre-heated water bath at 37 °C until only
a little lump of ice is le.
5. Pour tissue pieces onto the MACS SmartStrainer (70 µm).
6. Wash cryogenic vial with 1 mL of cell culture medium and
pour it onto the MACS SmartStrainer (70 µm).
7. Wash tissue pieces with additional 15 mL of cell culture
medium.
8. Pieces are ready to be used for downstream application, e.g.,
tissue dissociation using the MACS Tissue Dissociation Kits
in combination with the gentleMACS Octo Dissociator with
Heaters.
9. (Optional) To recover cells remaining in the medium,
centrifuge the medium for 5 minutes at 300×g.
10. (Optional) Aspirate supernatant and resuspend cell pellet in
appropriate buer according to downsteam application.
Refer to www.miltenyibiotec.com for all data sheets and protocols.
Miltenyi Biotec provides technical support worldwide. Visit
www.miltenyibiotec.com for local Miltenyi Biotec Technical
Support contact information.
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in the absence thereof, ONE (1) YEAR from date of delivery (“Product Warranty”).
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DEPENDING ON THE SPECIFIC APPLICATION. THE CUSTOMER IS SOLELY
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