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ATCC recommends that appropriate safety procedures be used when handling all primary cells and cell lines, especially those derived from human or other primate material. Detailed discussions of laboratory safety procedures are provided in Laboratory Safety: Principles and Practice, 2nd ed. (ASM Press, Washington, DC) (Fleming et al., 1995) and Caputo, J.L. Biosafety procedures in cell culture. (1988) J. Tissue Culture Methods 11:223. Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.] 

Human Material Precaution 

All tissues used for isolation are obtained under informed consent and conform to HIPAA standards to protect the privacy of the donor’s personal health information. It is best to use caution when handling any human cells. We recommend that all human cells be accorded the same level of biosafety consideration as cells known to carry HIV. With infectious virus assays or viral antigen assays, even a negative test result may leave open the possible existence of a latent viral genome.

The cells (HUtSMCs) are cryopreserved in the second passage to ensure the highest viability and plating efficiency. 

Vascular Cell Basal Medium (ATCC PCS-100-030), when supplemented with Vascular Cell Growth Kit (ATCC PCS-100-042), provides an ideal system to propagate uterine smooth muscle cells.

1. Obtain one vial of Primary Uterine Smooth Muscle Cells; Normal, Human (ATCC PCS-460-011) from the freezer? make sure that the caps of all components are tight.
2. Thaw the components of the growth kit (ATCC PCS-100-042) just prior to adding them to the basal medium (ATCC PCS-100-030).
3. Obtain one bottle of Vascular Cell Basal Medium (485 mL; PCS-100-030) from cold storage.
4. Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
5. Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component, as indicated in Table 1, to the bottle of basal medium using a separate sterile pipette for each transfer.

Table 1. Vascular Smooth Muscle Cell Growth Kit Components