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What Is the Name of the Area in Tissue Where Adult Stem Cells Are Found?

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Marrow-Derived Mesenchymal Stem Cells; Pattern, Human

PCS-500-012™

One ampul of Marrow-Derived Mesenchymal Stem Cells; Normal, Human (ATCC PCS-500-012™) containing a minimum of 1 x 106 viable cells (provided).

Product category

Human cells

Being

Homo man, human

Cell type
mesenchymal stem cell
Sound structure
mandril wrought, fibroblast-like
Tissue
Bone
Disease

Normal

Applications

Radica cubicle research

Product format
Frozen
Storage conditions

Vaporization phase of liquid nitrogen

Documentation

Biosafety Icon BSL 1

ATCC determines the biosafety level of a reincarnate supported our risk assessment equally radio-controlled by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Anthropomorphous Services. It is your responsibility to understand the hazards associated with the material per your organization's policies and procedures every bit well as any other applicable regulations equally implemented by your section operating room national agencies.

All tissues used for closing off are obtained under informed consent and conform to HIPAA regulations to protect the secrecy of the conferrer's Personally Identifiable Information. It is best to role caution when handling whatsoever fallible cells. We recommend that all fallible cells be accorded the same level of biosafety consideration as cells known to carry Human immunodeficiency virus (HIV) and other bloodborne pathogens. With infectious computer virus assays or viral antigen assays, even a negative test result may not throw out the possible action of the universe of a latent viral genome or catching micro-organism particles below the lower bound of detection of that essay.

ATCC recommends that set aside safety procedures be used when handling all primary cells and cell lines, especially those derived from human operating theater other archpriest fabric. Grip as a possibly biohazardous material victimization universal precautions. Cells derived from primate lymphoid tissue May fall under the regulations of 29 CFR 1910.1030 Bloodborne Pathogens.

ATCC extremely recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in musical nitrogen, IT is important to note that some vials may leak when submersed in liquid atomic number 7 and will slowly filling with liquid nitrogen. Upon melt, the transition of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its pileus with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures comprise stored in the vapor phase of liquid nitrogen preferably than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and take up been confirmed as in force in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be moved.

General

Specific applications
Adult stem cell specialization, regenerative medicine, mobile phone therapy, tissue engineering, multiplication of iPS cell lines

Characteristics

Cells per vial
≥ 1.0 x 106
Loudness
1.0 millilitre
Increment properties
Adherent
Age
adult
Ethnicity
Lot-specific
Gender
Lot-taxonomic category
Comments
The cells are cryopreserved in the second transit to ensure the highest viability and plating efficiency.

Mesenchymal Stem Cellular telephone Basal Mass medium, when supplemented with Mesenchymal Bow Cell Growth Kit out – for Bone up Marrow Derived MSCs, provides an apotheosis cell system to propagate mesenchymal stem cells. When maintained under best emergence conditions, ATCC Normal Human Debone Marrow-Derived Mesenchymal Stem Cells have been shown to be multipotent, subject of differentiating down the adipogenic, osteogenic and chondrogenic lineages.

Handling entropy

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry chalk publicity and immediately place the cells at a temperature downstairs ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
  1. Find one Mesenchymal Stem Cell Growth for Bone Sum-derived MSCs (ATCC PCS-500-041) from the freezer; make sure that the caps of altogether components are tight.
  2. Thaw the components of the ontogenesis kit just prior to adding them to the basal medium (ATCC PCS-500-030).
  3. Obtain one bottle of Mesenchymal Stem Cell Basal Medium (485 c) from cold storage.
  4. Decontaminate the external surfaces of altogether ontogenesis kit up component vials and the basal medium bottle past spraying them with 70% ethanol.
  5. Using aseptic technique and temporary in a laminar fall hood or biosafety locker, transfer the indicated volume of each development kit component, as indicated below, to the feeding bottle of basal medium using a separate sterile pipette for each transport.
    1. Marrow-Mesenchymal Stem Cell Growing Kit Components
      • FBS, 35 mL (Final concentration 7%)
      • rh IGF-1, 0.5 mL (Final engrossment 15 ng/mil)
      • Rhesus factor FGF-b, 0.5 mL (Final assiduousness 125 pg/c)
      • L-Alanyl-L-Glutamine, 6 mL (Final concentration 2.4 millimetre)

      2. Antimicrobials and phenol red are not required for proliferation merely English hawthorn be added if desired. The recommended volume of either of the facultative components to be added to the complete growth media is summarized downstairs.
    2. Addition of Antimicrobials/Antimycotics and Phenol Carmine (Optional):
      • Gentamicin-Amphotericin B Solvent, 0.5 cubic centimeter (Final density Gentamicin: 10 µg/mL, Amphotericin B: 25 nanogram/mL)
      • Penicillin-Streptomycin-Amphotericin B Solution, 0.5 mL (Final concentration Penicillin: 10 Units/mL, Streptomycin: 10 µg/mL, Amphotericin B: 25 nanogram/mL).
      • Carbolic acid Red, 0.5 mL (Final concentration 33 µM)
  1. Tightly cap the feeding bottle of complete growth medium and purl the contents softly to assure a homogeneous solution. Do not shake forcefully to avoid foaming. Label and date the bottle.
  2. Complete growing media should Be stored in the dark at 2°C to 8°C (do non freeze). When stored nether these conditions, complete growth media is stable for 30 days.
Reagents for subculture
  1. DPBS (ATCC 30-2200)
  2. Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) containing 0.05% Trypsin and 0.02% EDTA.
  3. Trypsin Neutralizing Solution (ATCC PCS-999-004)
Differentiation media for BM-MSCs
  1. Adipocyte Differentiation Toolkit (ATCC PCS-500-053)
  2. Osteocyte Differentiation Toolkit (ATCC PCS-500-052)
  3. Chondrocyte Differentiation Toolkit (ATCC PCS-500-051)
Required media
One nursing bottle of Mesenchymal Stem Cadre Radical Medium for Adipose, Umbilical cord and Marrow-derived MSCs (ATCC PCS-500-030) plus one Mesenchymal Stem Cell Growth Kit for Bone Marrow-copied MSCs (ATCC PCS-500-041) that contains the following growth supplements: FBS, rh FGF basic, rh IGF-I, and L-Alanyl-L-Glutamine.
Optional media supplements
  1. Gentamicin-Amphotericin B Answer (ATCC PCS-999-025)
  2. Penicillin-Streptomycin-Amphotericin B Solution (ATCC PCS-999-002)
  3. Phenol Red (ATCC PCS-999-001)
Treatment procedure
  1. Refer to the Certification of Analysis for the total number of viable cells recovered from this lot of    ATCC PCS-500-012.
  2. Using the total number of viable cells, determine how a great deal skin-deep area lavatory comprise inoculated to achieve an first seeding density of 5,000 cells per centimetre2.
  3. Prepare the desired compounding of flasks. Add 5 mL of staring growth medium per 25 cm2 of skin-deep surface area. Place the flasks in a 37°C, 5% CO2, humidified brooder and allow the media to pre-equilibrate to temperature and pH for 30 minutes preceding to adding cells.
  4. While the culture flasks equilibrate, remove one vial of ATCC PCS-500-012 from storage and thaw the cells by gentle agitation in a 37°C weewe bath. To reduce the possibility of contaminant, keep the O-ring and crest out of the water. Thawing should be rapid (approximately 1 to 2 minutes).
  5. Remove the vial from the water bath as shortly as the contents are liquid, and decontaminate past dipping in or spray with 70% ethanol. All operations from this charge onward should comprise carried out under puritanical aseptic conditions.
  6. Add the appropriate volume of complete growth moderate [volume = (1 mL x number of flasks to be seeded) -1 mL] into a sterile conical thermionic tube. Using a sterile pipette, transmit the cells from the cryovial to the conical tube. Mildly pipette the cells to homogenize the reprieve. Do not centrifuge.
  7. Transfer 1.0 mL of the cadre suspension to each of the pre-equilibrated culture flasks prepared in steps 1 to 3 of Handling Procedure for Frozen Cells and Founding of Culture. Pipet several times, then cap and gently rock all flask to evenly distribute the cells.
  8. Place the seeded culture flasks in the brooder at 37°C, 5% CO2 atmosphere. Brood for at any rate 24 hours before processing the cells further.
Subculturing procedure
  1. Passage formula Feces-MSCs when the culture has reached approximately 80% confluence.
  2. Warm both the Trypsin-EDTA for First-string Cells (ATCC PCS-999-003) and the Trypsin Neutralizing Solution (ATCC PCS-999-004) to elbow room temperature prior to dissociation. Warm the complete outgrowth sensitive to 37°C prior to use with the cells.
  3. For from each one flaskful, carefully aspirate the spent media without disturbing the monolayer.
  4. Rinse the cell bed one time with 3 to 5 cubic centimetre D-PBS (ATCC 30-2200) to remove substance medium.
  5. Add prewarmed trypsin-EDTA solution (1 to 2 milliliter for every 25 cm2) to each flask.
  6. Gently rock each flask to ensure complete coverage of the trypsin-EDTA solution over the cells.
  7. Observe the cells under the microscope. When the cells pull off from to each one other and round up (typically within 1 to 3 minutes), remove the flask from the microscope and gently hydrant it from several sides to encourage detachment of the cells from the flaskful surface.
  8. When the legal age of cells appear to have detached, quickly add an equal bulk of the Trypsin Neutralizing Resolution (ATCC PCS-999-004) to for each one flask. Mildly pipette Oregon purl the culture to ensure all of the trypsin-EDTA solution has been neutralized.
  9. Remove the dissociated cells to a sterile centrifuge tube and set aside while processing any remaining cells in the culture flask.
  10. Add 3 to 5 mL D-PBS (ATCC 30-2200) to the tissue refinement flaskful to collect any additional cells that power have been left behind.
  11. Transfer the cell/D-PBS suspension to the centrifuge tubing containing the trypsin-EDTA dissociated cells.
  12. Repeat steps 10 and 11 as needed until all cells have been collected from the flaskful.
  13. Centrifuge the cells at 270 x g for 5 minutes.
  14. Aspirate neutralized dissociation solution from the cell pellet and resuspend the cells in 2 to 8 mL fresh, prewarmed, over development medium.
  15. Count the cells and seed newborn culture flasks at a density of 5,000 viable cells per cm2.
  16. Place recently seedless flasks in a 37°C, 5% CO2 incubator for at least 24 to 48 hours before processing the cells advance. Bring up to Maintenance for guidelines along feeding.
Culture criminal maintenance
  1. Before beginning, prewarm complete growth media in a 37°C water bath. This wish take 'tween 10 and 30 minutes, contingent the volume. If using a lowercase volume of spiritualist (50 millilitre or less), warm only the bulk requisite in a sterile conelike tube. Avoid heating complete outgrowth media multiple times.
  2. 24 hours after seeding, remove the cells from the incubator and watch each flask under the microscope to determine percentage cellular merging.
  3. Carefully remove the gone media without disturbing the monolayer.
  4. Add 5 mL of fresh, prewarmed utter growth medium per 25 centimetre2 of surface region and recall the flasks to the incubator.
  5. Later 24 to 48 hours, view each flask under the microscope to determine percent cancellous confluence. If not ready to passage, repeat steps 3 and 4 as described above. When cultures have reached more or less 80% concourse, and are actively proliferating (many another mitotic figures are visible), IT is clip to subculture.
Note: BM-MSCs are contact inhibited. It is essential that the cells be subcultured Earlier reach 100% merging as postconfluent cells exhibit changes in morphology, slower proliferation, and reduced differentiation capacity after passaging.

Quality see specifications

Bacterial and fungal examination
Non detected
Mycoplasma taint
Not detected
Computer virus testing

HIV (HIV): Non detected

Hepatitis C computer virus (HCV): Not detected

Serum hepatitis virus (HBV): Not perceived

Population doubling capacity
≥ 10 in complete growth intermediate and digest specialization
Viability
≥ 70% when thawed from cryopreservation
Specific staining
Positive locution for CD29, CD44, CD73, CD90, CD105, and CD166
Perverse expression for CD14, CD34, CD19, and CD45

Legal disclaimers

Intended habituate
This product is intended for laboratory research use only. It is not intended for any animal or imperfect therapeutic use, any human or quail-like consumption, or any diagnostic practice.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of dispatch, provided that the customer has stored and handled the product according to the information included on the merchandise information sheet, website, and Certificate of Analysis. For livelihood cultures, ATCC lists the media conceptualisation and reagents that throw been found to embody good for the product. While else unspecified media and reagents may too produce o.k. results, a change in the ATCC and/or depositor-recommended protocols may affect the convalescence, growth, and/or function of the product. If an alternative medium formulation or reagent is victimized, the ATCC warranty for viability is no more valid.  Except Eastern Samoa expressly set forth herein, no different warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for science laborator research use only. It is not intended for any scorpion-like operating theater human therapeutic use, any human or animal wasting disease, operating theater any diagnostic use of goods and services. Whatever planned commercial use is prohibited without a license from ATCC.

Patc ATCC uses levelheaded efforts to let in true and improving-to-see information happening this product sheet, ATCC makes no warranties or representations arsenic to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does non warrant that such information has been confirmed to cost accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of whatever so much information.

This product is sent on the status that the customer is trusty for and assumes all risk and obligation in connection with the receipt, handling, storage, administration, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health operating theatre environmental endangerment. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC ware and whatever progeny Oregon modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'Eastern Samoa IS' with none representations or warranties whatsoever except equally expressly expound herein and in none effect shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, uncommon, incidental, or of import damages of any kindhearted in link with surgery arising out of the client's use of the product. While commonsense exploit is made to ensure genuineness and reliability of materials connected deposit, ATCC is not liable for restitution arising from the misidentification operating room misrepresentation of such materials.

Delight see the material transfer agreement (MTA) for promote inside information regarding the wont of this production. The MTA is visible at www.atcc.org.

References

Curated Citations

Al Madhoun A, et al. With chemicals Delimited Conditions Mediate an Efficient Initiation of Mesodermal Ancestry from Human Point Cord- and Marrow- Mesenchymal Stem Cells and Dental Pulp Pluripotent-Like Stem Cells. Cell Reprogram 20(1):9-16, 2018. PubMed: 29412734

Zheng B, et atomic number 13. Quantitative Magnetic Particle Imaging Monitors the Organ transplant, Biodistribution, and Clearance of Stem Cells In Vivo. Theranostics 6(3):291-301, 2016. PubMed: 26909106

Wang M, et al. Cold atmospheric plasma (Crownwork) surface nanomodified 3D printed polylactic acid (PLA) scaffoldsfor bone re-formation. Acta Biomater 46:256-265, 2016. PubMed: 27667017

Liu CJ, et al. Suppression of IL-8-Src signalling axis by 17β-estradiol inhibits quality mesenchymal stem cells-mediated gastric cancer invasion. J Cell Mol Med 20(5):962-72, 2016. PubMed: 26945908

Khalil S, et alii. A Cost-Effective Method to Assemble Biomimetic 3D Cell Culture Platforms. PLoS One 11(12):e0167116, 2016. PubMed: 27935982

Look at All Curated Citations for this Product

What Is the Name of the Area in Tissue Where Adult Stem Cells Are Found?

Source: https://www.atcc.org/products/pcs-500-012

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