Cell Types: Superficial Cells

Prepared by Kurt Schaberg
Cervical Cytology
Cell types Superficial Cells:
Small, pyknotic nucleus
Abundant cytoplasm (Often pink, can be blue)
Polygonal shape
Indicate abundant Estrogen
Intermediate Cells:
Abundant blue cytoplasm, polygonal shape
Larger, round to oval nuclei
Finer, normochromatic nuclei
→ Nuclei are important reference size
Basal/Parabasal Cell:
Minimal cytoplasm
Round to oval nuclei
Fine, but slightly dark chromatin
Usually few in number, unless atrophic
Endocervical Cells:
Uniform, Columnar cells
Polar, with round nucleus at one end
Majority of cytoplasm occupied by mucin
Arranged in flat sheets→ think “Honeycomb
Arranged in linear strips→ “Palisaded”
Endometrial Cells:
Small, High N:C ratio cells (almost all nucleus!)
Nucleus about the same size as an intermediate cell nucleus
Round nuclei with smooth chromatin, possible micronucleoli
Can be in large groups with outside epithelium and in inside stroma
Normal finding in first half of menstrual cycle if premenopausal
(Report if >50 yrs old)
Pap Smear Adequacy Criteria

(In practice, the number of cells is
Minimum number of well-visualized squamous cells for adequacy estimated based on sample photos,
Liquid-based preparation: 5,000 cells (ThinPrep and SurePath) and the cells aren’t counted).
Conventional Preparation: 8,000 to 12,000 cells
If obscuring elements cover >75% of epithelial cells → Unsatisfactory
Quality indicator: Presence of ≥10 endocervical cells or squamous metaplastic cells (reported, but not
required to be satisfactory for evaluation)
Any specimen with abnormal cells is considered adequate and should be reported!
Low-Grade Squamous Intraepithelial Lesion (LSIL)
Mature Keratinocytes (with lots of cytoplasm) AND:
• Enlarged nuclei (>3x normal intermediate cells)
• Nuclear membrane irregularities
• Hyperchromasia (“Rasinoid”)
• NO nucleoli
Optional:
• Perinuclear Halos = Koilocytes
• Large, irregular clearing
• Thick borders, like it was drawn with a
calligraphy pen
• Multinucleation
Caused by High and Low-risk HPV

May regress spontaneously!
Some findings, but “not enough”?

Consider “Atypical Squamous Cells of Undetermined
Significance” (ASCUS)
Can be either Quantitative (i.e., only rare atypical cells)
or Qualitative (e.g., only 2x nuclear enlargement)
High-Grade Squamous Intraepithelial Lesion (HSIL)

Immature keratinocytes (minimal cytoplasm, High N/C ratios) with:
• Markedly irregular nuclear contours
• (Hint: think in 3-dimensions)
• Look like boulders with all the irregularities
• Irregular chromatin and/or Hyperchromasia
Some findings, but “not enough”?

Consider “Atypical Squamous Cells—Cannot exclude HSIL” (ASC-H)
Can be either Quantitative (i.e., only rare atypical cells) or
Qualitative (e.g., only moderate atypia)
Squamous Cell Carcinoma
Non-keratinizing SCC may look like HSIL (similar findings)
Clues to invasion: “Tumor diathesis” (Necrotic debris)

Prominent nucleoli
Keratinizing SCC: Pleomorphic cells with hyperchromatic, irregular nuclei,

prominent orangeophilic (keratinizing) cytoplasm, and bizarre shapes (like
“Tadpoles” or snakes)
Tadpole Cell
Glandular Abnormalities
Reactive Endocervical Cells

Nuclear enlargement (4-5x),
Hyperchromasia, BUT round nuclei with smooth contours and
N:C ratios maintained. Prominent nucleoli.
Not too crowded. Mitoses, but no apoptosis.
Can see tubal metaplasia→ look for cilia!
For AIS, think “Feathery,”
like a bird’s wing.
Endocervical Adenocarcinoma In Situ (AIS):
Nuclei enlargement and crowding (cigar-like, think GI adenoma)
High N:C ratios with coarse, dark chromatin.
Cellular crowding with rosettes and “feathery edges”
Mitoses and apoptosis. No nucleoli.
Most strongly associated with HPV18 subtype
Adenocarcinoma
Variable, depending on site of origin/type.
Generally, more pleomorphic/irregular.
Endometrial cell nuclei larger than intermediate cell.
Features suggesting invasion: 1)Macronucleoli, 2)Tumor

diathesis, 3)increased single cells, and 4)irregular chromatin
Factors favoring endometrial adenocarcinoma (vs endocervix):

Neutrophils, less cytoplasm, smaller nuclei
Practically speaking, often diagnose as simply: “Atypical

Glandular Cells” using Bethesda System unless very
pleomorphic
Squamous metaplasia
Thick, “Dense” cytoplasm (consistent, dark teal)
Sharply defined cell borders.
Round, usually central nuclei
Normal nuclear size
Count as sampling of transition zone
Reparative/Inflammatory Changes
Classic “Repair”
Enlarged nuclei with Prominent Nucleoli.
Round nuclear contours with fine, pale chromatin.
Normal N:C ratios, but variably sized
Cohesive flat sheets of cells with “streaming” like pulled taffy
Background inflammation
General inflammatory change

Mild nuclear enlargement (<2x size)
Fine, pale chromatin
Often nucleoli
Can see small perinuclear clearing, but smaller, and more
even than koilocytic halos
Atrophy
Seen in LOW estrogen states:
Postmenopausal
Postpartum
Premenarche
Turner syndrome
Predominance of basal and parabasal cells (High N:C ratios)

Prone to injury→ often inflammation
Immature cells that can mimic HSIL, BUT:

Finely granular chromatin
Smooth nuclear contours
Can see “Blue blobs” (see →) and granular debris (mimicking

tumor diathesis, but no karyorrhectic nuclear debris)
Radiation Changes
Cytomegaly: Big cells with Big
nuclei (proportion N:C
maintained)
Large, bizarre cells
Cytoplasmic vacuolation and

polychromasia
Multinucleation
Follicular Cervicitis
Abundant lymphocytes (small rim of cytoplasm
around round nucleus, unlike HSIL, which is irregular)
Numerous tingible body macrophages
Variably sized lymphocytes with plasma cells.
IUD-Effect
Two characteristic findings:
1) Cells with abundant vacuolated cytoplasm
(mimicking Adenocarcinoma)
2) Cells with small, dark nuclei and scant

cytoplasm (mimicking HSIL)
Always know history! 1 2
Herpes
3 M’s
Molding of nuclei
Multinucleation
Margination of chromatin
“Ground glass” chromatin with eosinophilic nuclear inclusions
Can treat with acyclovir

“Shift” in flora
“Clue Cells” = squamous cells covered with shaggy bacilli and mixed
bacteria rather than the normal lactobacilli
Thick, milky vaginal discharge with a foul “fishy” odor

(positive “whiff” test after adding KOH)
Most commonly attributed to Gardenerella vaginalis.
Treat if symptomatic.
Trichomonas Vaginalis
Pear-shaped protozoan STD
Pale, eccentric elongate nucleus
Red cytoplasmic granules
Often associated with Leptothrix (non-pathogenic

filamentous bacterium).
Often acute inflammation and inflammatory halos

Treat with antibiotics
Candida
Fungal species that can cause infectious
throughout the GYN tract (and other areas).
Thick, “cottage cheese” discharge
Eosinophilic yeast forms and pseudohyphae and
hyphae (“Spaghetti and meatballs”)
Often tangled or skewering squamous cells
Can have variable associated inflammation or
inflammatory halos
Usually only treat if symptomatic
Actinomyces
Gram-positive anaerobic bacteria
Commonly associated with IUD (or other foreign body)
Long, filamentous organisms

Tangled clumps of bacteria that look like “cotton balls”
or “dust bunnies”
No need to remove IUD or treat if asymptomatic

General Management
This is a very generalized and abbreviated (but hopefully still helpful) summarization!
For a full review of current management guidelines, please refer to the ASCCP website:
http://www.asccp.org/guidelines
Pap result → Next step

Unsatisfactory → Repeat Pap in 2-4 months
NILM, but HPV positive (> 30 yrs) → Repeat co-testing at 1 yr, OR, HPV subtyping, if high-risk→ colpo
ASCUS → HPV testing (“if you Ask Us, you should get an HPV test”), if positive → colpo; Neg→ routine
LSIL → Colposcopy, unless HPV negative, then can do repeat co-testing at 1 yr
21-24 yo with ASCUS or LSIL → follow-up in 12 months (likely to clear spontaneously)
ASC-H → Colposcopy
HSIL→ Colposcopy or LEEP
Atypical glandular cells → Colposcopy with endocervical sampling and endometrial sampling
Screening Recommendations (according to the USPSTF):

Women 21-65 Cytology alone every 3 years
OR Women 30-65 Co-testing (cytology + HPV testing) every 5 yrs
(Don’t do HPV testing if less than 30 as high rate of positivity, but also clearance)
(Don’t do any testing before age 21, after 65 if they have had good prior screening, or after a
hysterectomy for benign reasons)
Human Papilloma Virus (HPV)

Sexually Transmitted Disease.
Circular double stranded DNA virus
Infects transformation zone→ establishes itself and replicates in basal cells
Detected in the vast majority of cervical cancers.
Viral genes responsible for transforming host cells by integrating into the host DNA and disrupting tumor
suppressor genes.
E6→ inactivates p53 (blocks apoptosis)
E7→ inactivates Rb (gets rid of cell cycle arrest→ uncontrolled growth)
However, many infections are naturally cleared, so only a minority persist and lead to cancer.
HPV subtypes
High-risk (associated with cervical cancer and HSIL, but can also cause LSIL)
16, 18, 31, 33
Type 16 is most commonly detected in cervical cancers
Type 18 is associated with Endocervical Adenocarcinoma
Low-risk (Not associated with HSIL. Instead associated with LSIL/condylomas)

6, 11, 42, 43

Cell Types: Superficial Cells

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Cell Types: Superficial Cells

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Prepared by Kurt Schaberg

Pap Smear Adequacy Criteria

Caused by High and Low-risk HPV

Some findings, but “not enough”?

High-Grade Squamous Intraepithelial Lesion (HSIL)

Some findings, but “not enough”?

Clues to invasion: “Tumor diathesis” (Necrotic debris)

Keratinizing SCC: Pleomorphic cells with hyperchromatic, irregular nuclei,

Reactive Endocervical Cells

Features suggesting invasion: 1)Macronucleoli, 2)Tumor

Factors favoring endometrial adenocarcinoma (vs endocervix):

Practically speaking, often diagnose as simply: “Atypical

Count as sampling of transition zone

General inflammatory change

Predominance of basal and parabasal cells (High N:C ratios)

Immature cells that can mimic HSIL, BUT:

Can see “Blue blobs” (see →) and granular debris (mimicking

Large, bizarre cells

Cytoplasmic vacuolation and

Numerous tingible body macrophages

Variably sized lymphocytes with plasma cells.

2) Cells with small, dark nuclei and scant

Always know history! 1 2

“Ground glass” chromatin with eosinophilic nuclear inclusions

Can treat with acyclovir

Thick, milky vaginal discharge with a foul “fishy” odor

Most commonly attributed to Gardenerella vaginalis.

Often associated with Leptothrix (non-pathogenic

Often acute inflammation and inflammatory halos

Long, filamentous organisms

No need to remove IUD or treat if asymptomatic

Pap result → Next step

Screening Recommendations (according to the USPSTF):

Human Papilloma Virus (HPV)

Low-risk (Not associated with HSIL. Instead associated with LSIL/condylomas)

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